e coli dh5α  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Thermo Fisher e coli dh5α
    Confirmation of the intracellular location of ChiA74∆s p in Escherichia coli and Bacillus thuringiensis 4Q7 using a chimeric construct with the green fluorescent protein (GFP) gene. (A) Schematic illustration that shows the fusion of chiA74 ∆s p with gfp . Panel 1, two constructs were developed, under regulation of the chiA74 wildtype promoter (wp) or under control of the cytA - p/ STAB-SD system. Lollipop indicates transcriptional terminator. Oligonucleotides used to make chimeric construct with gfp are shown in Table 1 . Panel 2, confirmation of chimeric constructs by PCR, primers used for amplification are showed in parenthesis and in Table 1 . L, 1 kb (kilobase) DNA Ladder (New England Biolabs); Lane 1, pEHchiA74∆ sp - gfp (primers: chiA74-1, chiA74-3); lane 2, pEHchiA74∆ sp - gfp (primers: chiA74-1, gfp-3); lane 3, pEHchiA74∆ sp - gfp (primers: gfp-1, chiA74-4); lane 4, pEB chi A74∆sp- gfp (primers: cytSTAB-1, chiA74-4); Lane 5, pEB chi A74∆sp- gfp (primers: cytSTAB-1, gfp-3); lane 6, pEB chi A74∆sp- gfp (primers: gfp-1, chiA74-4). (B) Phase contrast (left) and fluorescence micrographs (right) of recombinant strains of E. coli and B. thuringiensis strain 4Q7 expressing the chimeric protein ChiA74∆sp-GFP. Panel 1, E. coli- pEHchiA74∆ sp - gfp ; panel 2, 4Q7 - pEHchiA74∆ sp - gfp ; panel 3, E. coli- pEBchiA74∆ sp - gfp ; panel 4, 4Q7 - pEBchiA74∆ sp - gfp ; panel 5, E. coli <t>DH5α;</t> panel 6, 4Q7. Samples of recombinant strains of B. thuringiensis were collected at ~ 72 h.
    E Coli Dh5α, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 170 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli dh5α/product/Thermo Fisher
    Average 94 stars, based on 170 article reviews
    Price from $9.99 to $1999.99
    e coli dh5α - by Bioz Stars, 2020-07
    94/100 stars

    Images

    1) Product Images from "Bacillus thuringiensis subsp. kurstaki HD1 as a factory to synthesize alkali-labile ChiA74?sp chitinase inclusions, Cry crystals and spores for applied use"

    Article Title: Bacillus thuringiensis subsp. kurstaki HD1 as a factory to synthesize alkali-labile ChiA74?sp chitinase inclusions, Cry crystals and spores for applied use

    Journal: Microbial Cell Factories

    doi: 10.1186/1475-2859-13-15

    Confirmation of the intracellular location of ChiA74∆s p in Escherichia coli and Bacillus thuringiensis 4Q7 using a chimeric construct with the green fluorescent protein (GFP) gene. (A) Schematic illustration that shows the fusion of chiA74 ∆s p with gfp . Panel 1, two constructs were developed, under regulation of the chiA74 wildtype promoter (wp) or under control of the cytA - p/ STAB-SD system. Lollipop indicates transcriptional terminator. Oligonucleotides used to make chimeric construct with gfp are shown in Table 1 . Panel 2, confirmation of chimeric constructs by PCR, primers used for amplification are showed in parenthesis and in Table 1 . L, 1 kb (kilobase) DNA Ladder (New England Biolabs); Lane 1, pEHchiA74∆ sp - gfp (primers: chiA74-1, chiA74-3); lane 2, pEHchiA74∆ sp - gfp (primers: chiA74-1, gfp-3); lane 3, pEHchiA74∆ sp - gfp (primers: gfp-1, chiA74-4); lane 4, pEB chi A74∆sp- gfp (primers: cytSTAB-1, chiA74-4); Lane 5, pEB chi A74∆sp- gfp (primers: cytSTAB-1, gfp-3); lane 6, pEB chi A74∆sp- gfp (primers: gfp-1, chiA74-4). (B) Phase contrast (left) and fluorescence micrographs (right) of recombinant strains of E. coli and B. thuringiensis strain 4Q7 expressing the chimeric protein ChiA74∆sp-GFP. Panel 1, E. coli- pEHchiA74∆ sp - gfp ; panel 2, 4Q7 - pEHchiA74∆ sp - gfp ; panel 3, E. coli- pEBchiA74∆ sp - gfp ; panel 4, 4Q7 - pEBchiA74∆ sp - gfp ; panel 5, E. coli DH5α; panel 6, 4Q7. Samples of recombinant strains of B. thuringiensis were collected at ~ 72 h.
    Figure Legend Snippet: Confirmation of the intracellular location of ChiA74∆s p in Escherichia coli and Bacillus thuringiensis 4Q7 using a chimeric construct with the green fluorescent protein (GFP) gene. (A) Schematic illustration that shows the fusion of chiA74 ∆s p with gfp . Panel 1, two constructs were developed, under regulation of the chiA74 wildtype promoter (wp) or under control of the cytA - p/ STAB-SD system. Lollipop indicates transcriptional terminator. Oligonucleotides used to make chimeric construct with gfp are shown in Table 1 . Panel 2, confirmation of chimeric constructs by PCR, primers used for amplification are showed in parenthesis and in Table 1 . L, 1 kb (kilobase) DNA Ladder (New England Biolabs); Lane 1, pEHchiA74∆ sp - gfp (primers: chiA74-1, chiA74-3); lane 2, pEHchiA74∆ sp - gfp (primers: chiA74-1, gfp-3); lane 3, pEHchiA74∆ sp - gfp (primers: gfp-1, chiA74-4); lane 4, pEB chi A74∆sp- gfp (primers: cytSTAB-1, chiA74-4); Lane 5, pEB chi A74∆sp- gfp (primers: cytSTAB-1, gfp-3); lane 6, pEB chi A74∆sp- gfp (primers: gfp-1, chiA74-4). (B) Phase contrast (left) and fluorescence micrographs (right) of recombinant strains of E. coli and B. thuringiensis strain 4Q7 expressing the chimeric protein ChiA74∆sp-GFP. Panel 1, E. coli- pEHchiA74∆ sp - gfp ; panel 2, 4Q7 - pEHchiA74∆ sp - gfp ; panel 3, E. coli- pEBchiA74∆ sp - gfp ; panel 4, 4Q7 - pEBchiA74∆ sp - gfp ; panel 5, E. coli DH5α; panel 6, 4Q7. Samples of recombinant strains of B. thuringiensis were collected at ~ 72 h.

    Techniques Used: Construct, Polymerase Chain Reaction, Amplification, Fluorescence, Recombinant, Expressing

    Expression of ChiA74Δsp in Escherichia coli DH5α and Bacillus thuringiensis 4Q7. (A) Schematic illustration of the strategy used to delete the secretion signal peptide-encoding sequence (shown as a rectangle inside the open reading frame) of chiA74 to generate chiA74∆ s p. Two constructs were developed, the first under regulation of wildtype promoter (wp) and the second under control of the strong cytA-p /STAB-SD promoter system. Lollipop indicates the putative transcriptional terminator site. Oligonucleotide sequences used to delete the signal peptide-coding sequence are shown in Table 1 . (B) Evaluation of endochitinase activity using solubilized intracellular proteins produced in E. coli. Panel 1: Zymogram using 4-MU-(GlcNAc) 3 for detection. Lane 1, E. coli- pEHchiA74∆s p ; lane 2, without sample; lane 3, E. coli- pEBchiA74∆s p; lane 4, E. coli DH5α . Black arrow indicates the position of ChiA74∆s p in recombinant E. coli strains . Panel 2: (a) E. coli- pEHchiA74∆s p , (b) E. coli- pEBchiA74∆s p. No endochitinase activity was observed with E. coli DH5α . (C) Phase contrast microscopy of recombinant strains of B. thuringiensis . Panel 1, 4Q7; panel 2, 4Q7 - pEHchiA74∆s p ; panel 3, 4Q7 - pEBchiA74∆s p . Black arrows indicate the positions of ChiA74∆s p inclusions. (D) Endochitinase activity determined using solubilized intracellular proteins of recombinant strains of B. thuringiensis. Panel 1: lane 1, 4Q7; lane 2, 4Q7 - pEHchiA74∆s p ; lane 3, 4Q7 - pEBchiA74∆s p. Black arrow indicates the position of ChiA74∆s p in recombinant 4Q7 strains . Panel 2: (a) 4Q7 - pEHchiA74∆s p , (b) 4Q7 - pEBchiA74∆s p . Activity of recombinant bacteria was normalized with the residual intracellular endochitinase activity of 4Q7.
    Figure Legend Snippet: Expression of ChiA74Δsp in Escherichia coli DH5α and Bacillus thuringiensis 4Q7. (A) Schematic illustration of the strategy used to delete the secretion signal peptide-encoding sequence (shown as a rectangle inside the open reading frame) of chiA74 to generate chiA74∆ s p. Two constructs were developed, the first under regulation of wildtype promoter (wp) and the second under control of the strong cytA-p /STAB-SD promoter system. Lollipop indicates the putative transcriptional terminator site. Oligonucleotide sequences used to delete the signal peptide-coding sequence are shown in Table 1 . (B) Evaluation of endochitinase activity using solubilized intracellular proteins produced in E. coli. Panel 1: Zymogram using 4-MU-(GlcNAc) 3 for detection. Lane 1, E. coli- pEHchiA74∆s p ; lane 2, without sample; lane 3, E. coli- pEBchiA74∆s p; lane 4, E. coli DH5α . Black arrow indicates the position of ChiA74∆s p in recombinant E. coli strains . Panel 2: (a) E. coli- pEHchiA74∆s p , (b) E. coli- pEBchiA74∆s p. No endochitinase activity was observed with E. coli DH5α . (C) Phase contrast microscopy of recombinant strains of B. thuringiensis . Panel 1, 4Q7; panel 2, 4Q7 - pEHchiA74∆s p ; panel 3, 4Q7 - pEBchiA74∆s p . Black arrows indicate the positions of ChiA74∆s p inclusions. (D) Endochitinase activity determined using solubilized intracellular proteins of recombinant strains of B. thuringiensis. Panel 1: lane 1, 4Q7; lane 2, 4Q7 - pEHchiA74∆s p ; lane 3, 4Q7 - pEBchiA74∆s p. Black arrow indicates the position of ChiA74∆s p in recombinant 4Q7 strains . Panel 2: (a) 4Q7 - pEHchiA74∆s p , (b) 4Q7 - pEBchiA74∆s p . Activity of recombinant bacteria was normalized with the residual intracellular endochitinase activity of 4Q7.

    Techniques Used: Expressing, Sequencing, Construct, Activity Assay, Produced, Recombinant, Microscopy

    2) Product Images from "Natural microbial communities supporting the transfer of the IncP-1β plasmid pB10 exhibit a higher initial content of plasmids from the same incompatibility group"

    Article Title: Natural microbial communities supporting the transfer of the IncP-1β plasmid pB10 exhibit a higher initial content of plasmids from the same incompatibility group

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2014.00637

    Stability of plasmid pB10 and Escherichia coli DH5α in three river sediment microcosms (A–C) . Numbers indicate rate of disappearance (expressed in log per day) calculated from an exponential curve fit of the data (dotted lines). Each data point is an average of three values obtained from triplicate qPCR reactions, error bars represent standard deviation. Part of the data were presented previously in Bonot and Merlin (2010) .
    Figure Legend Snippet: Stability of plasmid pB10 and Escherichia coli DH5α in three river sediment microcosms (A–C) . Numbers indicate rate of disappearance (expressed in log per day) calculated from an exponential curve fit of the data (dotted lines). Each data point is an average of three values obtained from triplicate qPCR reactions, error bars represent standard deviation. Part of the data were presented previously in Bonot and Merlin (2010) .

    Techniques Used: Plasmid Preparation, Real-time Polymerase Chain Reaction, Standard Deviation

    Dissemination of plasmid pB10 (A) and relative abundance of IncP-1α/β plasmids (B) in various environmental communities maintained in microcosms. The ability of each environmental matrix to support the dissemination of plasmid pB10 was deduced from the comparison of pB10 and E. coli DH5α DNA relative stabilities in microcosms overtime (rate of disappearance), as exemplified in Figure 1 for sediments A–C. Transfer of pB10 was considered as effective (+) when pB10 stability appeared significantly greater than the stability of its initial donor host. Close rate of disappearance for both pB10 and E. coli DH5α reflects an absence of transfer (-). Values between brackets represent the number of the corresponding WWTPs. effluents [#1] were sampled twice in May 2011(#1A) and February 2012 (#1B). Each data point is an average of 3 values obtained from triplicate qPCR, where error bars represent standard deviation.
    Figure Legend Snippet: Dissemination of plasmid pB10 (A) and relative abundance of IncP-1α/β plasmids (B) in various environmental communities maintained in microcosms. The ability of each environmental matrix to support the dissemination of plasmid pB10 was deduced from the comparison of pB10 and E. coli DH5α DNA relative stabilities in microcosms overtime (rate of disappearance), as exemplified in Figure 1 for sediments A–C. Transfer of pB10 was considered as effective (+) when pB10 stability appeared significantly greater than the stability of its initial donor host. Close rate of disappearance for both pB10 and E. coli DH5α reflects an absence of transfer (-). Values between brackets represent the number of the corresponding WWTPs. effluents [#1] were sampled twice in May 2011(#1A) and February 2012 (#1B). Each data point is an average of 3 values obtained from triplicate qPCR, where error bars represent standard deviation.

    Techniques Used: Plasmid Preparation, Real-time Polymerase Chain Reaction, Standard Deviation

    3) Product Images from "Functions of the Mismatch Repair Gene mutS from Acinetobacter sp. Strain ADP1 †"

    Article Title: Functions of the Mismatch Repair Gene mutS from Acinetobacter sp. Strain ADP1 †

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.183.23.6822-6831.2001

    Cloning of mutS and flanking DNA from the chromosome of Acinetobacter sp. strain ADP1. Small arrows indicate the degenerate primers MUTSF2 (F) and MUTSR3 (R) used for PCR amplification of a 1.9-kb segment of mutS . Shaded regions indicate this portion of mutS throughout the figure. Strain ADP7003 was formed by integration of pZR7000 into the chromosome of strain ADP1. pGEM-3Zf(+) does not replicate in strain ADP1, so selection for ampicillin resistance (ap r ), encoded on the vector, demanded strain ADP7003. Digestion of chromosomal DNA from strain ADP7003 with Eco RI yielded a fragment containing pGEM-3Zf(+) fused to a segment of upstream DNA that included the 5′ end of mutS , and chromosomal DNA extending to the first Eco RI site upstream of the gene. Circularization of the restriction fragments by ligation followed by transformation into E. coli DH5α and selection for Ap r resulted in pZR7009. The 3′ end of mutS and downstream DNA were cloned in the same manner except that ADP7003 DNA was digested with Bam HI rather than Eco RI, and the resulting plasmid was designated pZR1010.
    Figure Legend Snippet: Cloning of mutS and flanking DNA from the chromosome of Acinetobacter sp. strain ADP1. Small arrows indicate the degenerate primers MUTSF2 (F) and MUTSR3 (R) used for PCR amplification of a 1.9-kb segment of mutS . Shaded regions indicate this portion of mutS throughout the figure. Strain ADP7003 was formed by integration of pZR7000 into the chromosome of strain ADP1. pGEM-3Zf(+) does not replicate in strain ADP1, so selection for ampicillin resistance (ap r ), encoded on the vector, demanded strain ADP7003. Digestion of chromosomal DNA from strain ADP7003 with Eco RI yielded a fragment containing pGEM-3Zf(+) fused to a segment of upstream DNA that included the 5′ end of mutS , and chromosomal DNA extending to the first Eco RI site upstream of the gene. Circularization of the restriction fragments by ligation followed by transformation into E. coli DH5α and selection for Ap r resulted in pZR7009. The 3′ end of mutS and downstream DNA were cloned in the same manner except that ADP7003 DNA was digested with Bam HI rather than Eco RI, and the resulting plasmid was designated pZR1010.

    Techniques Used: Clone Assay, Polymerase Chain Reaction, Amplification, Selection, Plasmid Preparation, Ligation, Transformation Assay

    4) Product Images from "Requirement of MrpH for Mannose-Resistant Proteus-Like Fimbria-Mediated Hemagglutination by Proteus mirabilis"

    Article Title: Requirement of MrpH for Mannose-Resistant Proteus-Like Fimbria-Mediated Hemagglutination by Proteus mirabilis

    Journal: Infection and Immunity

    doi:

    Western blot of partially purified fimbrial preparations of various E. coli DH5α strains. Partially purified fimbrial preparations (see Materials and Methods for details) were separated on an SDS–12.5% polyacrylamide gel and subjected to Western blot analysis with polyclonal rabbit antiserum raised against MrpA.
    Figure Legend Snippet: Western blot of partially purified fimbrial preparations of various E. coli DH5α strains. Partially purified fimbrial preparations (see Materials and Methods for details) were separated on an SDS–12.5% polyacrylamide gel and subjected to Western blot analysis with polyclonal rabbit antiserum raised against MrpA.

    Techniques Used: Western Blot, Purification

    Bacterial aggregation and MRHA patterns of the Δ mrpH mutant expressed in E. coli . (A) Pictures of 72-h static cultures of E. coli DH5α containing various constructs grown in 5 ml of Luria-Bertani broth at 37°C. The tubes were handled carefully to avoid disrupting the pellicle. (B) MRHA assay of the bacterial cultures in panel A were performed as described in Materials and Methods.
    Figure Legend Snippet: Bacterial aggregation and MRHA patterns of the Δ mrpH mutant expressed in E. coli . (A) Pictures of 72-h static cultures of E. coli DH5α containing various constructs grown in 5 ml of Luria-Bertani broth at 37°C. The tubes were handled carefully to avoid disrupting the pellicle. (B) MRHA assay of the bacterial cultures in panel A were performed as described in Materials and Methods.

    Techniques Used: Mutagenesis, Construct

    5) Product Images from "Maturation of molybdoenzymes and its influence on the pathogenesis of non-typeable Haemophilus influenzae"

    Article Title: Maturation of molybdoenzymes and its influence on the pathogenesis of non-typeable Haemophilus influenzae

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2015.01219

    Neutrophil-dependent killing of HI2019 strains and E. coli Dh5α . Neutrophils (2 * 10 ∧6 cells/mL) were incubated with 1:10 HI2019 WT , HI2019 Δ mobA , and HI2019 Δ mobA _comp strains and after 2 h of incubation, viable bacteria were measured by CFUs counting as described in Materials and Methods. Values shown are % viable bacteria at 2 h compared to the initial inoculum. E. coli Dh5α cells were used as a control.
    Figure Legend Snippet: Neutrophil-dependent killing of HI2019 strains and E. coli Dh5α . Neutrophils (2 * 10 ∧6 cells/mL) were incubated with 1:10 HI2019 WT , HI2019 Δ mobA , and HI2019 Δ mobA _comp strains and after 2 h of incubation, viable bacteria were measured by CFUs counting as described in Materials and Methods. Values shown are % viable bacteria at 2 h compared to the initial inoculum. E. coli Dh5α cells were used as a control.

    Techniques Used: Incubation

    6) Product Images from "Improving Acetate Tolerance of Escherichia coli by Rewiring Its Global Regulator cAMP Receptor Protein (CRP)"

    Article Title: Improving Acetate Tolerance of Escherichia coli by Rewiring Its Global Regulator cAMP Receptor Protein (CRP)

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0077422

    Growth profile. Mutant strain A2, the control (DH5α Δ crp harbouring plasmid pKCP containing native crp operon) and DH5α Δ crp strain harbouring blank plasmid pKC in M9 medium at 37 °C ( A ) 0 g/L sodium acetate ( B ) 10 g/L sodium acetate ( C ) 15 g/L sodium acetate. Average values and standard deviations were calculated from triplicate experiments.
    Figure Legend Snippet: Growth profile. Mutant strain A2, the control (DH5α Δ crp harbouring plasmid pKCP containing native crp operon) and DH5α Δ crp strain harbouring blank plasmid pKC in M9 medium at 37 °C ( A ) 0 g/L sodium acetate ( B ) 10 g/L sodium acetate ( C ) 15 g/L sodium acetate. Average values and standard deviations were calculated from triplicate experiments.

    Techniques Used: Mutagenesis, Plasmid Preparation

    Cell growth of E. coli DH5α overexpressing uxaB in M9 medium, with E. coli DH5α harbouring pCA24N as control. ( A ) 0 g/L sodium acetate ( B ) 5 g/L sodium acetate. Average values and standard deviations were calculated from triplicate experiments.
    Figure Legend Snippet: Cell growth of E. coli DH5α overexpressing uxaB in M9 medium, with E. coli DH5α harbouring pCA24N as control. ( A ) 0 g/L sodium acetate ( B ) 5 g/L sodium acetate. Average values and standard deviations were calculated from triplicate experiments.

    Techniques Used:

    7) Product Images from "Amino acid residues 201-205 in C-terminal acidic tail region plays a crucial role in antibacterial activity of HMGB1"

    Article Title: Amino acid residues 201-205 in C-terminal acidic tail region plays a crucial role in antibacterial activity of HMGB1

    Journal: Journal of Biomedical Science

    doi: 10.1186/1423-0127-16-83

    Western blot analysis of the recombination proteins . Identification of the expression of the His-tagged recombinant proteins in E. Coli DH5α by Western blot. 1, DHFR; 2, rHMGB1 A box; 3, rHMGB1 B box; 4, rHMGB1; 5, tHMGB1; 6, mHMGB1 -211-215; 7, mHMGB1- 206-215; 8, mHMGB1 -201-215; 9, mHMGB1 -196-215; 10, mHMGB1 -191-215; 11, mHMGB1 -186-200; 12, mHMGB1 -196-210; 13, mHMGB1 -196-205; 14, mHMGB1 -198-207; 15, mHMGB1 -201-210; 16, mHMGB1 -201-205.
    Figure Legend Snippet: Western blot analysis of the recombination proteins . Identification of the expression of the His-tagged recombinant proteins in E. Coli DH5α by Western blot. 1, DHFR; 2, rHMGB1 A box; 3, rHMGB1 B box; 4, rHMGB1; 5, tHMGB1; 6, mHMGB1 -211-215; 7, mHMGB1- 206-215; 8, mHMGB1 -201-215; 9, mHMGB1 -196-215; 10, mHMGB1 -191-215; 11, mHMGB1 -186-200; 12, mHMGB1 -196-210; 13, mHMGB1 -196-205; 14, mHMGB1 -198-207; 15, mHMGB1 -201-210; 16, mHMGB1 -201-205.

    Techniques Used: Western Blot, Expressing, Recombinant

    Antibacterial activity assay of eleven different deleted mutants lacking different several amino acid residues in C-terminal acidic tail region of HMGB1 with the method of dilution in the test tubes . The antibacterial activities of eleven different deleted mutants lacking different several amino acid residues in C-terminal acidic tail region of HMGB1 were compared by the method of dilution in the test tubes. The bacteria used in this experiment included SA, JM109, ATCC 25922, DH5α and PA. Each experiment was performed in triplicate and repeated three times. DHFR protein purified by the same system was used as a negative control. Data showed as mean ± standard deviation and were analyzed with One-Way ANOVA. * P
    Figure Legend Snippet: Antibacterial activity assay of eleven different deleted mutants lacking different several amino acid residues in C-terminal acidic tail region of HMGB1 with the method of dilution in the test tubes . The antibacterial activities of eleven different deleted mutants lacking different several amino acid residues in C-terminal acidic tail region of HMGB1 were compared by the method of dilution in the test tubes. The bacteria used in this experiment included SA, JM109, ATCC 25922, DH5α and PA. Each experiment was performed in triplicate and repeated three times. DHFR protein purified by the same system was used as a negative control. Data showed as mean ± standard deviation and were analyzed with One-Way ANOVA. * P

    Techniques Used: Activity Assay, Purification, Negative Control, Standard Deviation

    Antibacterial activity assay of rHMGB1, tHMGB1, rHMGB1 A box, B box, and C peptide with the method of dilution in the test tubes . The antibacterial activities of rHMGB1, tHMGB1, rHMGB1 A box, B box, and C peptide were compared by the method of dilution in the test tubes as described in the Materials and Methods . The bacteria used in this experiment included SA, JM109, ATCC 25922, DH5α and PA. Each experiment was performed in triplicate and repeated three times. DHFR protein purified by the same system was used as a negative control. Data showed as mean ± standard deviation and were analyzed with One-Way ANOVA. * P
    Figure Legend Snippet: Antibacterial activity assay of rHMGB1, tHMGB1, rHMGB1 A box, B box, and C peptide with the method of dilution in the test tubes . The antibacterial activities of rHMGB1, tHMGB1, rHMGB1 A box, B box, and C peptide were compared by the method of dilution in the test tubes as described in the Materials and Methods . The bacteria used in this experiment included SA, JM109, ATCC 25922, DH5α and PA. Each experiment was performed in triplicate and repeated three times. DHFR protein purified by the same system was used as a negative control. Data showed as mean ± standard deviation and were analyzed with One-Way ANOVA. * P

    Techniques Used: Activity Assay, Purification, Negative Control, Standard Deviation

    8) Product Images from "Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants"

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants

    Journal:

    doi: 10.1128/AEM.71.11.7224-7228.2005

    Growth of E. coli DH5α strains expressing tyrA fbr genes in the presence of m -fluoro- d , l -tyrosine. Eight strains of E. coli DH5α harboring different plasmids were monitored: — , pZE21:: tyrA WT ; + , pZE21:: tyrA fbr-5 ; ▪,
    Figure Legend Snippet: Growth of E. coli DH5α strains expressing tyrA fbr genes in the presence of m -fluoro- d , l -tyrosine. Eight strains of E. coli DH5α harboring different plasmids were monitored: — , pZE21:: tyrA WT ; + , pZE21:: tyrA fbr-5 ; ▪,

    Techniques Used: Expressing

    9) Product Images from "A Novel Epimerase That Converts GlcNAc-P-P-undecaprenol to GalNAc-P-P-undecaprenol in Escherichia coli O157 *"

    Article Title: A Novel Epimerase That Converts GlcNAc-P-P-undecaprenol to GalNAc-P-P-undecaprenol in Escherichia coli O157 *

    Journal:

    doi: 10.1074/jbc.M109.061630

    Z3206 does not complement glycosylation of AcrA in a Gne-dependent glycosylation system. Periplasmic extracts prepared from E. coli DH5α cells carrying the AcrA expression plasmid and the pgl operon Δ gne complemented with pMLBAD:Z3206
    Figure Legend Snippet: Z3206 does not complement glycosylation of AcrA in a Gne-dependent glycosylation system. Periplasmic extracts prepared from E. coli DH5α cells carrying the AcrA expression plasmid and the pgl operon Δ gne complemented with pMLBAD:Z3206

    Techniques Used: Expressing, Plasmid Preparation

    10) Product Images from "Defining the Growth Conditions and Promoter-Proximal DNA Sequences Required for Activation of Gene Expression by CreBC in Escherichia coli "

    Article Title: Defining the Growth Conditions and Promoter-Proximal DNA Sequences Required for Activation of Gene Expression by CreBC in Escherichia coli

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.00108-08

    Effect of disruption of genes of the cre locus on creD expression. Expression was measured using β-galactosidase reporter plasmid pUB6070. DH5α (wild type [WT]), DH5αΔ creA (A), DH5αΔ creB (B), DH5αΔ
    Figure Legend Snippet: Effect of disruption of genes of the cre locus on creD expression. Expression was measured using β-galactosidase reporter plasmid pUB6070. DH5α (wild type [WT]), DH5αΔ creA (A), DH5αΔ creB (B), DH5αΔ

    Techniques Used: Expressing, Plasmid Preparation

    11) Product Images from "Development of a Sensitive and Specific Enzyme-Linked Immunosorbent Assay for Detecting and Quantifying CMY-2 and SHV ?-Lactamases"

    Article Title: Development of a Sensitive and Specific Enzyme-Linked Immunosorbent Assay for Detecting and Quantifying CMY-2 and SHV ?-Lactamases

    Journal: Journal of Clinical Microbiology

    doi: 10.1128/JCM.40.6.1947-1957.2002

    Immunoblotting. (a) Immunoblot of various amounts of purified CMY-2 β-lactamase probed with 1 μg of anti-CMY-2 antibody/ml. (b) Immunoblot of various amounts of purified SHV-1 β-lactamase probed with 1 μg of anti-SHV-1 antibody/ml. (c) Immunoblot of various β-lactamase-producing strains probed with 1 μg of anti-CMY-2 antibody/ml. Strains, listed from left to right, included E coli DH10B carrying plasmid pBC SK(−) with the SHV-1 β-lactamase, strains producing K-1 and ACT-1 β-lactamases, strain DH5α/pUC18 producing the TEM-1 β-lactamase, a cefepime-resistant E. aerogenes strain producing a β-lactamase (EA), and a strain expressing the P99 Amp C β-lactamase; in addition, E. coli J53-2-derived strains 194-61 and 194 and E. coli strain 20 (EC20) are clinical and laboratory strains producing CMY-2 β-lactamase. (d) Identical immunoblots of strains E. coli DH10B/pUC18 producing TEM-1 β-lactamase, E. coli DH10B/pBC SK(−) producing SHV-1 β-lactamase, and E. coli J53-2-derived 194-61 producing CMY-2 β-lactamase probed with anti-TEM antibody (1:100 dilution) or 1 μg of anti-SHV antibody/ml.
    Figure Legend Snippet: Immunoblotting. (a) Immunoblot of various amounts of purified CMY-2 β-lactamase probed with 1 μg of anti-CMY-2 antibody/ml. (b) Immunoblot of various amounts of purified SHV-1 β-lactamase probed with 1 μg of anti-SHV-1 antibody/ml. (c) Immunoblot of various β-lactamase-producing strains probed with 1 μg of anti-CMY-2 antibody/ml. Strains, listed from left to right, included E coli DH10B carrying plasmid pBC SK(−) with the SHV-1 β-lactamase, strains producing K-1 and ACT-1 β-lactamases, strain DH5α/pUC18 producing the TEM-1 β-lactamase, a cefepime-resistant E. aerogenes strain producing a β-lactamase (EA), and a strain expressing the P99 Amp C β-lactamase; in addition, E. coli J53-2-derived strains 194-61 and 194 and E. coli strain 20 (EC20) are clinical and laboratory strains producing CMY-2 β-lactamase. (d) Identical immunoblots of strains E. coli DH10B/pUC18 producing TEM-1 β-lactamase, E. coli DH10B/pBC SK(−) producing SHV-1 β-lactamase, and E. coli J53-2-derived 194-61 producing CMY-2 β-lactamase probed with anti-TEM antibody (1:100 dilution) or 1 μg of anti-SHV antibody/ml.

    Techniques Used: Purification, Plasmid Preparation, Activated Clotting Time Assay, Transmission Electron Microscopy, Expressing, Derivative Assay, Western Blot

    12) Product Images from "Characterization of a minimal pKW2124 replicon from Weissella cibaria KLC140 and its application for the construction of the Weissella expression vector pKUCm1"

    Article Title: Characterization of a minimal pKW2124 replicon from Weissella cibaria KLC140 and its application for the construction of the Weissella expression vector pKUCm1

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2015.00035

    Evaluation of pKUCm series and pMR1 vector re-transformation efficiencies from W. confusa ATCC 10881 into the same strain (black scale bar), compared with the vectors prepared from Escherichia coli DH5α into W. confusa ATCC 10881 (gray scale bar). Re-transformation of the vectors from Weissella improved the transformation efficiency.
    Figure Legend Snippet: Evaluation of pKUCm series and pMR1 vector re-transformation efficiencies from W. confusa ATCC 10881 into the same strain (black scale bar), compared with the vectors prepared from Escherichia coli DH5α into W. confusa ATCC 10881 (gray scale bar). Re-transformation of the vectors from Weissella improved the transformation efficiency.

    Techniques Used: Plasmid Preparation, Transformation Assay

    13) Product Images from "Anti-virulence properties of an antifreeze protein"

    Article Title: Anti-virulence properties of an antifreeze protein

    Journal: Cell reports

    doi: 10.1016/j.celrep.2014.09.034

    Binding of P1 to Staphylococcus aureus interferes with biofilm formation in vitro. A: S. aureus SA113, the methicillin-resistant USA300 JE2 isolate, L. monocytogenes EGDe and E. coli DH5α were incubated with biotinylated P1 (pos ctrl)in DMSO. Following washing, bound peptide was detected by immunoblot. DMSO or SA113 without peptide incubation served as negative controls. B: S. aureus SA113 and USA300 were preincubated in PBS supplemented with P1 or sP1 before incubation with GST-IAFGP. Following removal of unbound GST-IAFGP, GST-IAFGP associated with the bacterial pellet was detected by immunoblot. Recombinant GST-IAFGP was used as positive control for immunoblot. C: Biofilm formation of S. aureus SA113 and USA300 JE2 cultures supplemented with P1 or sP1 were measured using Safranin stains. Results are means ± SEM of 3 independent experiments performed in quadruplicate, representative images of the biofilm stain are shown (One-Way-ANOVA with Tukey’s post-test). Bacteria cultured in BHI/G (Medium) or supplemented with PBS serve as controls. D: PNAG formation of S. aureus supplemented with P1 or sP1 was visualized by immunoblot. Results are means ± SEM of 3 independent experiments, with representative images of the stained biofilm (One-Way-ANOVA with Tukey’s post-test). Bacteria cultured in BHI/G (Medium) without protein addition were used as reference.
    Figure Legend Snippet: Binding of P1 to Staphylococcus aureus interferes with biofilm formation in vitro. A: S. aureus SA113, the methicillin-resistant USA300 JE2 isolate, L. monocytogenes EGDe and E. coli DH5α were incubated with biotinylated P1 (pos ctrl)in DMSO. Following washing, bound peptide was detected by immunoblot. DMSO or SA113 without peptide incubation served as negative controls. B: S. aureus SA113 and USA300 were preincubated in PBS supplemented with P1 or sP1 before incubation with GST-IAFGP. Following removal of unbound GST-IAFGP, GST-IAFGP associated with the bacterial pellet was detected by immunoblot. Recombinant GST-IAFGP was used as positive control for immunoblot. C: Biofilm formation of S. aureus SA113 and USA300 JE2 cultures supplemented with P1 or sP1 were measured using Safranin stains. Results are means ± SEM of 3 independent experiments performed in quadruplicate, representative images of the biofilm stain are shown (One-Way-ANOVA with Tukey’s post-test). Bacteria cultured in BHI/G (Medium) or supplemented with PBS serve as controls. D: PNAG formation of S. aureus supplemented with P1 or sP1 was visualized by immunoblot. Results are means ± SEM of 3 independent experiments, with representative images of the stained biofilm (One-Way-ANOVA with Tukey’s post-test). Bacteria cultured in BHI/G (Medium) without protein addition were used as reference.

    Techniques Used: Binding Assay, In Vitro, Incubation, Recombinant, Positive Control, Staining, Cell Culture

    14) Product Images from "Importance of the GP Dipeptide of the “Antiporter Motif” and other Membrane-Embedded Proline and Glycine residues in Tetracycline Efflux Protein Tet(L)"

    Article Title: Importance of the GP Dipeptide of the “Antiporter Motif” and other Membrane-Embedded Proline and Glycine residues in Tetracycline Efflux Protein Tet(L)

    Journal: Biochemistry

    doi: 10.1021/bi050762c

    Transport assays of Tet(L) Gly 155 mutants in comparison with wild type Tet(L). Left panel: Uptake of [ 3 H]Tc-Co 2+ into everted vesicles from the indicated transformants of E. coli DH5α were carried out as described under Materials and Methods.
    Figure Legend Snippet: Transport assays of Tet(L) Gly 155 mutants in comparison with wild type Tet(L). Left panel: Uptake of [ 3 H]Tc-Co 2+ into everted vesicles from the indicated transformants of E. coli DH5α were carried out as described under Materials and Methods.

    Techniques Used:

    15) Product Images from "Association of Neisseria gonorrhoeae OpaCEA with Dendritic Cells Suppresses Their Ability to Elicit an HIV-1-Specific T Cell Memory Response"

    Article Title: Association of Neisseria gonorrhoeae OpaCEA with Dendritic Cells Suppresses Their Ability to Elicit an HIV-1-Specific T Cell Memory Response

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0056705

    Association of N. gonorrhoeae Opa CEA with MDDCs down-regulated CD83 expression, but did not affect IL-12 induction. A) A representative experiment from a healthy participant is shown. Immature MDDCs were infected with individual isogenic gonococcal strain or E.coli DH5α at an MOI of 10 in complete RPMI 1640 medium. Medium alone and CD40LT were included for negative and positive controls, respectively. MDDCs were harvested and expression of surface molecules was assayed by flow cytometric analysis. Values represent the mean flurorescence intensity subtracted from the value of matched isotype control mouse mAbs (shaded gray histogram). B) Summary data of CD83 expression on MDDCs surface obtained from all five participants of healthy blood donors are shown. Statistical comparisons of data pooled from five participants were performed between Opa CEA -expressing N309 and other gonococci strains, E.coli DH5α, or CD40LT treatment: N309 vs N302, p
    Figure Legend Snippet: Association of N. gonorrhoeae Opa CEA with MDDCs down-regulated CD83 expression, but did not affect IL-12 induction. A) A representative experiment from a healthy participant is shown. Immature MDDCs were infected with individual isogenic gonococcal strain or E.coli DH5α at an MOI of 10 in complete RPMI 1640 medium. Medium alone and CD40LT were included for negative and positive controls, respectively. MDDCs were harvested and expression of surface molecules was assayed by flow cytometric analysis. Values represent the mean flurorescence intensity subtracted from the value of matched isotype control mouse mAbs (shaded gray histogram). B) Summary data of CD83 expression on MDDCs surface obtained from all five participants of healthy blood donors are shown. Statistical comparisons of data pooled from five participants were performed between Opa CEA -expressing N309 and other gonococci strains, E.coli DH5α, or CD40LT treatment: N309 vs N302, p

    Techniques Used: Expressing, Infection, Flow Cytometry

    Association of N. gonorrhoeae Opa CEA with MDDCs decreased the sensitization of allogeneic mixed lymphocyte proliferations. Immature MDDCs infected, uninfected, or CD40LT-treated for 3 days were plated into 96-well round-bottom tissue-culture plates with allogeneic PBMCs at a ratio of 1:20 (MDDCs to T cells). Cells were cocultured for 3 days and pulsed with 2 µCi (0.076 Mbq) [ 3 H]thymidine for an additional 6 h. [ 3 H]Thymidine incorporation in harvested cells was assayed by liquid scintillation spectrometry. These data are mean ± SEM of triplicates and representative of four independent experiments with different donors. Statistical comparisons of data pooled from the five participants were performed between Opa CEA -expressing N309 and other gonococci strains, E.coli DH5α, or CD40LT treatment: N309 vs N302, p
    Figure Legend Snippet: Association of N. gonorrhoeae Opa CEA with MDDCs decreased the sensitization of allogeneic mixed lymphocyte proliferations. Immature MDDCs infected, uninfected, or CD40LT-treated for 3 days were plated into 96-well round-bottom tissue-culture plates with allogeneic PBMCs at a ratio of 1:20 (MDDCs to T cells). Cells were cocultured for 3 days and pulsed with 2 µCi (0.076 Mbq) [ 3 H]thymidine for an additional 6 h. [ 3 H]Thymidine incorporation in harvested cells was assayed by liquid scintillation spectrometry. These data are mean ± SEM of triplicates and representative of four independent experiments with different donors. Statistical comparisons of data pooled from the five participants were performed between Opa CEA -expressing N309 and other gonococci strains, E.coli DH5α, or CD40LT treatment: N309 vs N302, p

    Techniques Used: Infection, Expressing

    Effects of N. gonorrhoeae infection on HIV-1-specific CTL memory response. PBMCs from HIV-1-seropositive individuals were cocultured with HLA-restricted peptide-pulsed or nonpulsed autologous MDDCs that were previously infected for 72 h with N. gonorrhoeae N302, N303, N309, or N496, or E.coli DH5α, or CD40LT treatment at 1 µg/ml. On day 10 of coculture, HIV-1-specific CTL activity was assayed by intracellular staining and flow cytometric analysis of IFN-γ-producing CD8 + T cells. A) representative intracellular IFN-γ data obtained from Pt#1 of HIV-1 seropositive individual are shown. Cells were gated for CD3 and CD8 to enumerate IFN-γ-producing CD8 + T cells only. B) Summary data of intracellular IFN-γ production in CD8 + T cells from Pt#2 – Pt#5 are graphically depicted. C) Summary data of IFN-γ-producing CD8 + T cells from all patients tested are graphically depicted. The experiments from Pt#1 and Pt#2 were repeated with similar results. Statistical comparisons of data pooled from five participants were performed between Opa CEA -expressing N309 and other gonococci strains, E.coli DH5α, or CD40LT treatment: N309 vs N302, p
    Figure Legend Snippet: Effects of N. gonorrhoeae infection on HIV-1-specific CTL memory response. PBMCs from HIV-1-seropositive individuals were cocultured with HLA-restricted peptide-pulsed or nonpulsed autologous MDDCs that were previously infected for 72 h with N. gonorrhoeae N302, N303, N309, or N496, or E.coli DH5α, or CD40LT treatment at 1 µg/ml. On day 10 of coculture, HIV-1-specific CTL activity was assayed by intracellular staining and flow cytometric analysis of IFN-γ-producing CD8 + T cells. A) representative intracellular IFN-γ data obtained from Pt#1 of HIV-1 seropositive individual are shown. Cells were gated for CD3 and CD8 to enumerate IFN-γ-producing CD8 + T cells only. B) Summary data of intracellular IFN-γ production in CD8 + T cells from Pt#2 – Pt#5 are graphically depicted. C) Summary data of IFN-γ-producing CD8 + T cells from all patients tested are graphically depicted. The experiments from Pt#1 and Pt#2 were repeated with similar results. Statistical comparisons of data pooled from five participants were performed between Opa CEA -expressing N309 and other gonococci strains, E.coli DH5α, or CD40LT treatment: N309 vs N302, p

    Techniques Used: Infection, CTL Assay, Activity Assay, Staining, Flow Cytometry, Expressing

    Gonococcal binding and internalization to iMDDCs. A ) Immature MDDCs were incubated with FITC-labeled gonococcal strains N302, N303, N309, and N496, or E.coli DH5α bacteria at an MOI of 10 for 30 min at RT. Bacterial binding was determined by measuring the percentage of cells that bound FITC-labeled bacteria using flow cytometric analysis. The percentage of cells with bound FITC-labeled bacteria is indicated in each condition. B) Bacterial binding to iMDDCs is not mediated by DC-SIGN on the cell surface. Mannan blocking assays were carried out site-by-site with the bacterial binding tests. The percentage of cells with bound FITC-labeled bacteria is indicated in each condition in the presence of mannan at 20 µg/ml. C) Internalization of gonococcal strains and E.coli DH5α into iMDDCs. Immature MDDCs were allowed to adhere onto coverslips pre-coated with 0.2% gelatin. These cells were pulsed with gonococcal strains or E.coli DH5α (MOI = 100) prelabeled with Texas red-X-succinimidyl ester at 37 o C for 1 h. Extracellular bacteria were then labeled with the polyclonal anti-gonococcal serum, followed by a staining with a BODIPY-FL-conjugated secondary Ab. Immature MDDCs were then permeabilized with 0.4% Triton X-100 and stained with Phalloidin-FITC. Intracellular (red) versus extracellular (yellow) bacteria with MDDCs (green) were then distinguished by visualization with a Leica DM-IRBE inverted fluorescence microscope. The magnification for all conditions is 40.
    Figure Legend Snippet: Gonococcal binding and internalization to iMDDCs. A ) Immature MDDCs were incubated with FITC-labeled gonococcal strains N302, N303, N309, and N496, or E.coli DH5α bacteria at an MOI of 10 for 30 min at RT. Bacterial binding was determined by measuring the percentage of cells that bound FITC-labeled bacteria using flow cytometric analysis. The percentage of cells with bound FITC-labeled bacteria is indicated in each condition. B) Bacterial binding to iMDDCs is not mediated by DC-SIGN on the cell surface. Mannan blocking assays were carried out site-by-site with the bacterial binding tests. The percentage of cells with bound FITC-labeled bacteria is indicated in each condition in the presence of mannan at 20 µg/ml. C) Internalization of gonococcal strains and E.coli DH5α into iMDDCs. Immature MDDCs were allowed to adhere onto coverslips pre-coated with 0.2% gelatin. These cells were pulsed with gonococcal strains or E.coli DH5α (MOI = 100) prelabeled with Texas red-X-succinimidyl ester at 37 o C for 1 h. Extracellular bacteria were then labeled with the polyclonal anti-gonococcal serum, followed by a staining with a BODIPY-FL-conjugated secondary Ab. Immature MDDCs were then permeabilized with 0.4% Triton X-100 and stained with Phalloidin-FITC. Intracellular (red) versus extracellular (yellow) bacteria with MDDCs (green) were then distinguished by visualization with a Leica DM-IRBE inverted fluorescence microscope. The magnification for all conditions is 40.

    Techniques Used: Binding Assay, Incubation, Labeling, Flow Cytometry, Blocking Assay, Staining, Fluorescence, Microscopy

    16) Product Images from "Use of Chimeric Type IV Secretion Systems to Define Contributions of Outer Membrane Subassemblies for Contact-Dependent Translocation"

    Article Title: Use of Chimeric Type IV Secretion Systems to Define Contributions of Outer Membrane Subassemblies for Contact-Dependent Translocation

    Journal: Molecular microbiology

    doi: 10.1111/mmi.13700

    Chimeric T4SSs support conjugative DNA transfer and activate T6SS killing. A) Sequences encoding the outer membrane core complex (OMCC) subunits TraN, TraO, and the C-terminal half (residues 194–386) of TraF were replaced with corresponding genes or gene fragments from the A. tumefaciens VirB, E. coli R388 Trw, or B. pertussis Ptl systems on mini-pKM101 plasmid pCGR108. The chimeric T4SSs composed of the inner membrane complex (IMC) of pKM101 (yellow) joined to the OMCCs from the VirB, Trw, or Ptl systems (color-coded) are modeled on the R388 T4SS 3–10 ). B) E. coli donors carrying pCGR108 derivatives encoding the only the Tra pKM101 IMC or OMCC, or the IMC::OMCC chimeras transferred the mobilizable plasmid pJG42 at the frequencies shown in transconjugants per donor (Tc’s/D) in solid-surface (histogram, solid bars) or liquid (stippled bars) matings, and were resistant to IKe phage infection (S, sensitive; R, resistant). C) E. coli survival when cultivated in the absence or presence of P. aeruginosa PAO1. E. coli DH5α cells lacked or produced intact or variant forms of the Tra pKM101 T4SS depicted. Statistical significance is shown based on a Student’s t test corresponding to the values of plasmid-free DH5α or growth in the absence of P. aeruginosa (NS, not significant; * P
    Figure Legend Snippet: Chimeric T4SSs support conjugative DNA transfer and activate T6SS killing. A) Sequences encoding the outer membrane core complex (OMCC) subunits TraN, TraO, and the C-terminal half (residues 194–386) of TraF were replaced with corresponding genes or gene fragments from the A. tumefaciens VirB, E. coli R388 Trw, or B. pertussis Ptl systems on mini-pKM101 plasmid pCGR108. The chimeric T4SSs composed of the inner membrane complex (IMC) of pKM101 (yellow) joined to the OMCCs from the VirB, Trw, or Ptl systems (color-coded) are modeled on the R388 T4SS 3–10 ). B) E. coli donors carrying pCGR108 derivatives encoding the only the Tra pKM101 IMC or OMCC, or the IMC::OMCC chimeras transferred the mobilizable plasmid pJG42 at the frequencies shown in transconjugants per donor (Tc’s/D) in solid-surface (histogram, solid bars) or liquid (stippled bars) matings, and were resistant to IKe phage infection (S, sensitive; R, resistant). C) E. coli survival when cultivated in the absence or presence of P. aeruginosa PAO1. E. coli DH5α cells lacked or produced intact or variant forms of the Tra pKM101 T4SS depicted. Statistical significance is shown based on a Student’s t test corresponding to the values of plasmid-free DH5α or growth in the absence of P. aeruginosa (NS, not significant; * P

    Techniques Used: Plasmid Preparation, Infection, Produced, Variant Assay

    Requirements for activation of T6SS killing by P. aeruginosa PAO1. E. coli DH5α lacking or producing the Tra pKM101 T4SS composed of the His 6 -TraF variants shown; in each case, the traF allele was substituted for wild-type traF by incorporation into the pKM101 tra locus on plasmid pCGR108. Statistical significance is shown based on a Student’s t test corresponding to the values of plasmid-free DH5α or growth in the absence of P. aeruginosa (NS, not significant; * P
    Figure Legend Snippet: Requirements for activation of T6SS killing by P. aeruginosa PAO1. E. coli DH5α lacking or producing the Tra pKM101 T4SS composed of the His 6 -TraF variants shown; in each case, the traF allele was substituted for wild-type traF by incorporation into the pKM101 tra locus on plasmid pCGR108. Statistical significance is shown based on a Student’s t test corresponding to the values of plasmid-free DH5α or growth in the absence of P. aeruginosa (NS, not significant; * P

    Techniques Used: Activation Assay, Plasmid Preparation

    17) Product Images from "Maturation of molybdoenzymes and its influence on the pathogenesis of non-typeable Haemophilus influenzae"

    Article Title: Maturation of molybdoenzymes and its influence on the pathogenesis of non-typeable Haemophilus influenzae

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2015.01219

    Neutrophil-dependent killing of HI2019 strains and E. coli Dh5α . Neutrophils (2 * 10 ∧6 cells/mL) were incubated with 1:10 HI2019 WT , HI2019 Δ mobA , and HI2019 Δ mobA _comp strains and after 2 h of incubation, viable bacteria were measured by CFUs counting as described in Materials and Methods. Values shown are % viable bacteria at 2 h compared to the initial inoculum. E. coli Dh5α cells were used as a control.
    Figure Legend Snippet: Neutrophil-dependent killing of HI2019 strains and E. coli Dh5α . Neutrophils (2 * 10 ∧6 cells/mL) were incubated with 1:10 HI2019 WT , HI2019 Δ mobA , and HI2019 Δ mobA _comp strains and after 2 h of incubation, viable bacteria were measured by CFUs counting as described in Materials and Methods. Values shown are % viable bacteria at 2 h compared to the initial inoculum. E. coli Dh5α cells were used as a control.

    Techniques Used: Incubation

    18) Product Images from "Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants"

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants

    Journal:

    doi: 10.1128/AEM.71.11.7224-7228.2005

    Growth of E. coli DH5α strains expressing tyrA fbr genes in the presence of m -fluoro- d , l -tyrosine. Eight strains of E. coli DH5α harboring different plasmids were monitored: — , pZE21:: tyrA WT ; + , pZE21:: tyrA fbr-5 ; ▪,
    Figure Legend Snippet: Growth of E. coli DH5α strains expressing tyrA fbr genes in the presence of m -fluoro- d , l -tyrosine. Eight strains of E. coli DH5α harboring different plasmids were monitored: — , pZE21:: tyrA WT ; + , pZE21:: tyrA fbr-5 ; ▪,

    Techniques Used: Expressing

    19) Product Images from "Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants"

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants

    Journal:

    doi: 10.1128/AEM.71.11.7224-7228.2005

    Growth of E. coli DH5α strains expressing tyrA fbr genes in the presence of m -fluoro- d , l -tyrosine. Eight strains of E. coli DH5α harboring different plasmids were monitored: — , pZE21:: tyrA WT ; + , pZE21:: tyrA fbr-5 ; ▪,
    Figure Legend Snippet: Growth of E. coli DH5α strains expressing tyrA fbr genes in the presence of m -fluoro- d , l -tyrosine. Eight strains of E. coli DH5α harboring different plasmids were monitored: — , pZE21:: tyrA WT ; + , pZE21:: tyrA fbr-5 ; ▪,

    Techniques Used: Expressing

    20) Product Images from "Pseudomonas aeruginosa Elastase Provides an Escape from Phagocytosis by Degrading the Pulmonary Surfactant Protein-A"

    Article Title: Pseudomonas aeruginosa Elastase Provides an Escape from Phagocytosis by Degrading the Pulmonary Surfactant Protein-A

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0027091

    Δ lasB mutant bacteria are resistant to SP-A-mediated membrane permeabilization. Membrane permeabilization assays were performed with 1×10 8 of E. coli DH5α or P. aeruginosa exposed to hSP-A (50 µg/ml) for 120 min. Three independent experiments were performed in triplicates. The mean + standard deviation from one representative experiment is shown. The membrane permeabilization activity of hSP-A against PAO1, Δ lasB and PDO240lasB was not statistically different among all three P. aeruginosa strains. * p
    Figure Legend Snippet: Δ lasB mutant bacteria are resistant to SP-A-mediated membrane permeabilization. Membrane permeabilization assays were performed with 1×10 8 of E. coli DH5α or P. aeruginosa exposed to hSP-A (50 µg/ml) for 120 min. Three independent experiments were performed in triplicates. The mean + standard deviation from one representative experiment is shown. The membrane permeabilization activity of hSP-A against PAO1, Δ lasB and PDO240lasB was not statistically different among all three P. aeruginosa strains. * p

    Techniques Used: Mutagenesis, Standard Deviation, Activity Assay

    21) Product Images from "Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a – Escherichia coli DH5α system"

    Article Title: Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a – Escherichia coli DH5α system

    Journal: Veterinary World

    doi: 10.14202/vetworld.2018.557-561

    PCR amplification of E.coli DH5α clones Lane 2 to 7 –positive amplicons Lane1- DNA marker
    Figure Legend Snippet: PCR amplification of E.coli DH5α clones Lane 2 to 7 –positive amplicons Lane1- DNA marker

    Techniques Used: Polymerase Chain Reaction, Amplification, Clone Assay, Marker

    22) Product Images from "Transfer of Herb-Resistance Plasmid From Escherichia coli to Staphylococcus aureus Residing in the Human Urinary Tract"

    Article Title: Transfer of Herb-Resistance Plasmid From Escherichia coli to Staphylococcus aureus Residing in the Human Urinary Tract

    Journal: Jundishapur Journal of Microbiology

    doi: 10.5812/jjm.15056

    Resistance Plasmid Pattern Lane M, molecular size marker; lane D, plasmid fragment of E. coli CP9 as a donor; lane R1 and R2, trans conjugants of E. coli DH5α obtained using E. coli CP9 as a donor.
    Figure Legend Snippet: Resistance Plasmid Pattern Lane M, molecular size marker; lane D, plasmid fragment of E. coli CP9 as a donor; lane R1 and R2, trans conjugants of E. coli DH5α obtained using E. coli CP9 as a donor.

    Techniques Used: Plasmid Preparation, Marker

    23) Product Images from "What an Escherichia coli Mutant Can Teach Us About the Antibacterial Effect of Chlorophyllin"

    Article Title: What an Escherichia coli Mutant Can Teach Us About the Antibacterial Effect of Chlorophyllin

    Journal: Microorganisms

    doi: 10.3390/microorganisms7020059

    Influence of different chlorophyllin concentrations on the growth of ( A ) Escherichia coli DH5α, ( B ) Bacillus subtilis 168 and ( C ) Escherichia coli NR698 (initial cell number: ~10 8 cfu/mL). Bacteria were cultured for 24 h at 37 °C exposed to light (12 mW/cm 2 ; bright plots, upper row) or in darkness (grey plots, lower row). To determine the lower limit of efficacy, additional chlorophyllin concentrations were tested: ( D ) Escherichia coli DH5α, 20↑100 mg/L ( E ) Bacillus subtilis 168, 1.0↓0.01 mg/L and ( F ) Escherichia coli NR698, 1.0↓0.01 mg/L. Depicted are means (growth curves) together with corresponding 95% confidence limits (bar charts). Blue numbers indicate respective chlorophyllin concentrations. * p
    Figure Legend Snippet: Influence of different chlorophyllin concentrations on the growth of ( A ) Escherichia coli DH5α, ( B ) Bacillus subtilis 168 and ( C ) Escherichia coli NR698 (initial cell number: ~10 8 cfu/mL). Bacteria were cultured for 24 h at 37 °C exposed to light (12 mW/cm 2 ; bright plots, upper row) or in darkness (grey plots, lower row). To determine the lower limit of efficacy, additional chlorophyllin concentrations were tested: ( D ) Escherichia coli DH5α, 20↑100 mg/L ( E ) Bacillus subtilis 168, 1.0↓0.01 mg/L and ( F ) Escherichia coli NR698, 1.0↓0.01 mg/L. Depicted are means (growth curves) together with corresponding 95% confidence limits (bar charts). Blue numbers indicate respective chlorophyllin concentrations. * p

    Techniques Used: Cell Culture

    Effect of chlorophyllin on the early growth phase of ( A ) Escherichia coli DH5α, ( B ) Bacillus subtilis 168, and ( C ) Escherichia coli NR698. Liquid cultures of bacteria (initial cell number: 10 6 cfu/mL) were incubated in Erlenmeyer flasks in standard LB medium (red) and in presence of a chlorophyllin concentration of 22 mg/L (green). Cells grew either illuminated with 12 mW/cm 2 (bright plots, upper row) or in darkness (grey plots, lower row). Depicted are measured values (circles) and fitted curves (solid lines) with corresponding 95% confidence limits (red and green areas). Dashed lines describe growth of example cultures in LB medium + MeOH/KOH (solvent of chlorophyllin).
    Figure Legend Snippet: Effect of chlorophyllin on the early growth phase of ( A ) Escherichia coli DH5α, ( B ) Bacillus subtilis 168, and ( C ) Escherichia coli NR698. Liquid cultures of bacteria (initial cell number: 10 6 cfu/mL) were incubated in Erlenmeyer flasks in standard LB medium (red) and in presence of a chlorophyllin concentration of 22 mg/L (green). Cells grew either illuminated with 12 mW/cm 2 (bright plots, upper row) or in darkness (grey plots, lower row). Depicted are measured values (circles) and fitted curves (solid lines) with corresponding 95% confidence limits (red and green areas). Dashed lines describe growth of example cultures in LB medium + MeOH/KOH (solvent of chlorophyllin).

    Techniques Used: Incubation, Concentration Assay

    ( A ) Chemical structure of isolated chlorophyllin. ( B ) Alterations of chlorophyllin’s absorption spectrum in darkness and ( C ) exposed to light with 12 mW/cm 2 . ( D ) Reduction of chlorophyllin concentration exposed to different light intensities. ( E ) Required light exposure times to sterilize an E. coli DH5α suspension culture initial cell number: ~10 6 cfu/mL) with a light intensity of 12 mW/cm 2 using different chlorophyllin concentrations. ( F ) Required light exposure times to sterilize an E. coli DH5α suspension culture (initial cell number: ~10 6 cfu/mL) with a chlorophyllin concentration of 25 mg/L and different light intensities. For 12.5% light intensity no sterilization was observed within 300 min. Depicted are means ± standard deviation.
    Figure Legend Snippet: ( A ) Chemical structure of isolated chlorophyllin. ( B ) Alterations of chlorophyllin’s absorption spectrum in darkness and ( C ) exposed to light with 12 mW/cm 2 . ( D ) Reduction of chlorophyllin concentration exposed to different light intensities. ( E ) Required light exposure times to sterilize an E. coli DH5α suspension culture initial cell number: ~10 6 cfu/mL) with a light intensity of 12 mW/cm 2 using different chlorophyllin concentrations. ( F ) Required light exposure times to sterilize an E. coli DH5α suspension culture (initial cell number: ~10 6 cfu/mL) with a chlorophyllin concentration of 25 mg/L and different light intensities. For 12.5% light intensity no sterilization was observed within 300 min. Depicted are means ± standard deviation.

    Techniques Used: Isolation, Concentration Assay, Standard Deviation

    Schematic presentation of evaluation of CFU ability after incubation to chlorophyllin. Differently dense liquid cultures of ( A ) Escherichia coli DH5α, ( B ) Bacillus subtilis 168, and ( C ) Escherichia coli NR698 were supplemented with different chlorophyllin concentrations between 0.1 and 25 mg/L. Cells grew in 96-well matrix plates either illuminated with 12 mW/cm 2 ( A – C ) or protected from light ( D – F ). Samples (2.5 µL) were drawn at different time points and transferred onto LB agar plates. After overnight incubation at 37 °C in the dark, colony growth was analyzed. Dot size quantifies colony growth. Original pictures of the agar plates can be found in Figure S1 .
    Figure Legend Snippet: Schematic presentation of evaluation of CFU ability after incubation to chlorophyllin. Differently dense liquid cultures of ( A ) Escherichia coli DH5α, ( B ) Bacillus subtilis 168, and ( C ) Escherichia coli NR698 were supplemented with different chlorophyllin concentrations between 0.1 and 25 mg/L. Cells grew in 96-well matrix plates either illuminated with 12 mW/cm 2 ( A – C ) or protected from light ( D – F ). Samples (2.5 µL) were drawn at different time points and transferred onto LB agar plates. After overnight incubation at 37 °C in the dark, colony growth was analyzed. Dot size quantifies colony growth. Original pictures of the agar plates can be found in Figure S1 .

    Techniques Used: Incubation

    Fluorescence microscopic images of ( A ) Escherichia coli DH5α and ( B ) Escherichia coli NR698 after exposure to chlorophyllin (red fluorescence). Assumed modes of action of chlorophyllin against Gram-positive and Gram-negative bacteria. ( C ) Chlorophyllin (Chl) molecules cannot pass the intact outer membrane of Gram-negative bacteria. ( D ) Chlorophyllin is degraded in light. Degradation products can enter both Gram-negative and Gram-positive cells. The red crosses indicate cell death.
    Figure Legend Snippet: Fluorescence microscopic images of ( A ) Escherichia coli DH5α and ( B ) Escherichia coli NR698 after exposure to chlorophyllin (red fluorescence). Assumed modes of action of chlorophyllin against Gram-positive and Gram-negative bacteria. ( C ) Chlorophyllin (Chl) molecules cannot pass the intact outer membrane of Gram-negative bacteria. ( D ) Chlorophyllin is degraded in light. Degradation products can enter both Gram-negative and Gram-positive cells. The red crosses indicate cell death.

    Techniques Used: Fluorescence

    24) Product Images from "Stenotrophomonas maltophilia Encodes a VirB/VirD4 Type IV Secretion System That Modulates Apoptosis in Human Cells and Promotes Competition against Heterologous Bacteria, Including Pseudomonas aeruginosa"

    Article Title: Stenotrophomonas maltophilia Encodes a VirB/VirD4 Type IV Secretion System That Modulates Apoptosis in Human Cells and Promotes Competition against Heterologous Bacteria, Including Pseudomonas aeruginosa

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00457-19

    Survival of S. maltophilia wild-type and virB10 mutant strains when cocultured with heterologous bacteria. S. maltophilia (Sm) K279a (WT), virB10 mutant NUS15 ( virB10 ), or complemented mutant NUS15(p virB10 ) ( virB10/virB10+ ) was mixed with E. coli strain DH5α (Ec) (A), K. pneumoniae strain KPPR1 (Kp) (B), or P. aeruginosa strain 7700 (Pa) (C) at ratios of ∼200:1, 150:1, 100:1, and 50:1, spotted onto LB agar. After 24 h of incubation, the numbers of each strain were determined by plating serial dilutions of the bacterial growth area on selective media. Results are presented as the ratios of S. maltophilia CFU to the CFU of other species at t = 0 and t =24 h. The dagger symbols indicate when the ending ratio was
    Figure Legend Snippet: Survival of S. maltophilia wild-type and virB10 mutant strains when cocultured with heterologous bacteria. S. maltophilia (Sm) K279a (WT), virB10 mutant NUS15 ( virB10 ), or complemented mutant NUS15(p virB10 ) ( virB10/virB10+ ) was mixed with E. coli strain DH5α (Ec) (A), K. pneumoniae strain KPPR1 (Kp) (B), or P. aeruginosa strain 7700 (Pa) (C) at ratios of ∼200:1, 150:1, 100:1, and 50:1, spotted onto LB agar. After 24 h of incubation, the numbers of each strain were determined by plating serial dilutions of the bacterial growth area on selective media. Results are presented as the ratios of S. maltophilia CFU to the CFU of other species at t = 0 and t =24 h. The dagger symbols indicate when the ending ratio was

    Techniques Used: Mutagenesis, Incubation

    25) Product Images from "Chronic Prosthetic Hip Infection Caused by a Small-Colony Variant of Escherichia coli"

    Article Title: Chronic Prosthetic Hip Infection Caused by a Small-Colony Variant of Escherichia coli

    Journal: Journal of Clinical Microbiology

    doi:

    Immunoblotting. Cell lysates of E. coli DH5α and the two colony types of Z-2376 grown under aerobic or anaerobic conditions on TSA-blood agar plates were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to nitrocellulose sheets, and tested against antisera. (A) Antiserum from the patient diluted 1:500; (B) pooled antisera from 10 healthy volunteers diluted 1:100. Lanes 1, DH5α; lanes 2, large-colony type of Z-2376 grown aerobically; lanes 3, small-colony type of Z-2376 grown aerobically; lanes 4, large-colony type of Z-2376 grown anaerobically; lanes 5, small-colony type of Z-2376 grown anaerobically; lanes 6, prestained low-molecular-weight marker.
    Figure Legend Snippet: Immunoblotting. Cell lysates of E. coli DH5α and the two colony types of Z-2376 grown under aerobic or anaerobic conditions on TSA-blood agar plates were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to nitrocellulose sheets, and tested against antisera. (A) Antiserum from the patient diluted 1:500; (B) pooled antisera from 10 healthy volunteers diluted 1:100. Lanes 1, DH5α; lanes 2, large-colony type of Z-2376 grown aerobically; lanes 3, small-colony type of Z-2376 grown aerobically; lanes 4, large-colony type of Z-2376 grown anaerobically; lanes 5, small-colony type of Z-2376 grown anaerobically; lanes 6, prestained low-molecular-weight marker.

    Techniques Used: Polyacrylamide Gel Electrophoresis, Molecular Weight, Marker

    Southern hybridization. Two identical nylon membranes carrying immobilized, Eco RI-digested, genomic DNA were hybridized with the hemB probe (A) and the hemD probe (B). Lanes 1, DH5α; lanes 2: small-colony type of Z-2376; lanes 3, large-colony type of Z-2376; lanes M, marker ( Hin dIII-digested λ DNA). The localization of the different fragments at 23.7, 9.46, 6.66, 4.2, 2.25, and 1.96 kb (top to bottom) are indicated by lines.
    Figure Legend Snippet: Southern hybridization. Two identical nylon membranes carrying immobilized, Eco RI-digested, genomic DNA were hybridized with the hemB probe (A) and the hemD probe (B). Lanes 1, DH5α; lanes 2: small-colony type of Z-2376; lanes 3, large-colony type of Z-2376; lanes M, marker ( Hin dIII-digested λ DNA). The localization of the different fragments at 23.7, 9.46, 6.66, 4.2, 2.25, and 1.96 kb (top to bottom) are indicated by lines.

    Techniques Used: Hybridization, Marker

    26) Product Images from "Kinetic Properties of Four Plasmid-Mediated AmpC ?-Lactamases"

    Article Title: Kinetic Properties of Four Plasmid-Mediated AmpC ?-Lactamases

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.49.10.4240-4246.2005

    MICs of E. coli DH5α strains and overproduction of the plasmid-encoded class C β-lactamases.
    Figure Legend Snippet: MICs of E. coli DH5α strains and overproduction of the plasmid-encoded class C β-lactamases.

    Techniques Used: Plasmid Preparation

    27) Product Images from "Murine immunization with CS21 pili or LngA major subunit of enterotoxigenic Escherichia coli (ETEC) elicits systemic and mucosal immune responses and inhibits ETEC gut colonization"

    Article Title: Murine immunization with CS21 pili or LngA major subunit of enterotoxigenic Escherichia coli (ETEC) elicits systemic and mucosal immune responses and inhibits ETEC gut colonization

    Journal: Veterinary microbiology

    doi: 10.1016/j.vetmic.2016.02.001

    LngA-recombinant and CS21 pili antigens recognized by monoclonal anti-LngA and polyclonal murine sera Panel A. LngA-his tag recombinant and CS21purified native pili preparation were separated in an SDS-PAGE gel. The Coomassie blue stained gel is shown on the left and the Western blot labeled with anti-LngA monoclonal antibody is shown on the right. CS21 pili preparations were derived from E9034A ETEC strain; LngA-his tag protein was obtained from Genscript; BL21 E. coli was used as negative control; and E9034AΔ lngA pili preparation was used as LngA negative control. Panel B . Western blot of CS21 and CS8 pili from ETEC strains from different geographic locations using murine anti-LngA polyclonal sera. Purified CS21 and CS8 pili from ETEC strains originated in different countries were evaluated by Western blot using two different antisera, one derived from a mouse immunized IP with LngA-his tag antigen plus IFA and the second obtained from a mouse immunized IP with CS21 pili antigen plus IFA. Pili preparations from E. coli DH5α E. coli BL21 and CS21 + ETEC E9034A strains were used as negative and positive controls and LngA-His recombinant protein was used as LngA protein control. The size of LngA is 22 kDa.
    Figure Legend Snippet: LngA-recombinant and CS21 pili antigens recognized by monoclonal anti-LngA and polyclonal murine sera Panel A. LngA-his tag recombinant and CS21purified native pili preparation were separated in an SDS-PAGE gel. The Coomassie blue stained gel is shown on the left and the Western blot labeled with anti-LngA monoclonal antibody is shown on the right. CS21 pili preparations were derived from E9034A ETEC strain; LngA-his tag protein was obtained from Genscript; BL21 E. coli was used as negative control; and E9034AΔ lngA pili preparation was used as LngA negative control. Panel B . Western blot of CS21 and CS8 pili from ETEC strains from different geographic locations using murine anti-LngA polyclonal sera. Purified CS21 and CS8 pili from ETEC strains originated in different countries were evaluated by Western blot using two different antisera, one derived from a mouse immunized IP with LngA-his tag antigen plus IFA and the second obtained from a mouse immunized IP with CS21 pili antigen plus IFA. Pili preparations from E. coli DH5α E. coli BL21 and CS21 + ETEC E9034A strains were used as negative and positive controls and LngA-His recombinant protein was used as LngA protein control. The size of LngA is 22 kDa.

    Techniques Used: Recombinant, SDS Page, Staining, Western Blot, Labeling, Derivative Assay, Negative Control, Purification, Immunofluorescence

    28) Product Images from "An engineered autotransporter-based surface expression vector enables efficient display of Affibody molecules on OmpT-negative E. coli as well as protease-mediated secretion in OmpT-positive strains"

    Article Title: An engineered autotransporter-based surface expression vector enables efficient display of Affibody molecules on OmpT-negative E. coli as well as protease-mediated secretion in OmpT-positive strains

    Journal: Microbial Cell Factories

    doi: 10.1186/s12934-014-0179-z

    Evaluation of OmpT-mediated release and small-scale purification of displayed Affibody molecule. A . Flow-cytometric analysis for comparison of surface expression of recombinant proteins between p AraBAD -Z IgG -EC in OmpT-negative E. coli BL21 and p AraBAD -Z IgG -EC in OmpT-positive E. coli DH5α. B . SDS-PAGE showing IMAC-purified supernatants from p AraBAD -Z IgG -EC in OmpT-negative E. coli BL21 and p AraBAD -Z IgG -EC in OmpT-positive E. coli DH5α. Supernatants from non-induced samples, indicated with (−), were included as controls. C . Mass spectrum from ESI-MS analysis of IMAC-purified supernatants from p AraBAD -Z IgG -EC in OmpT-positive E. coli DH5α. Theoretical molecular weight of the OmpT-cleaved Z IgG -His 6 is 9712 Da. D . SPR-based biosensor analysis on the IMAC-purified Z IgG -His 6 . Response units (RU) on the y-axis and time on the x-axis. Representative sensorgrams from injection of Z IgG -His 6 at three different concentrations over human IgG immobilized on the chip surface. Injections were performed in duplicates.
    Figure Legend Snippet: Evaluation of OmpT-mediated release and small-scale purification of displayed Affibody molecule. A . Flow-cytometric analysis for comparison of surface expression of recombinant proteins between p AraBAD -Z IgG -EC in OmpT-negative E. coli BL21 and p AraBAD -Z IgG -EC in OmpT-positive E. coli DH5α. B . SDS-PAGE showing IMAC-purified supernatants from p AraBAD -Z IgG -EC in OmpT-negative E. coli BL21 and p AraBAD -Z IgG -EC in OmpT-positive E. coli DH5α. Supernatants from non-induced samples, indicated with (−), were included as controls. C . Mass spectrum from ESI-MS analysis of IMAC-purified supernatants from p AraBAD -Z IgG -EC in OmpT-positive E. coli DH5α. Theoretical molecular weight of the OmpT-cleaved Z IgG -His 6 is 9712 Da. D . SPR-based biosensor analysis on the IMAC-purified Z IgG -His 6 . Response units (RU) on the y-axis and time on the x-axis. Representative sensorgrams from injection of Z IgG -His 6 at three different concentrations over human IgG immobilized on the chip surface. Injections were performed in duplicates.

    Techniques Used: Purification, Flow Cytometry, Expressing, Recombinant, SDS Page, Mass Spectrometry, Molecular Weight, SPR Assay, Injection, Chromatin Immunoprecipitation

    29) Product Images from "Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a – Escherichia coli DH5α system"

    Article Title: Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a – Escherichia coli DH5α system

    Journal: Veterinary World

    doi: 10.14202/vetworld.2018.557-561

    PCR amplification of E.coli DH5α clones Lane 2 to 7 –positive amplicons Lane1- DNA marker
    Figure Legend Snippet: PCR amplification of E.coli DH5α clones Lane 2 to 7 –positive amplicons Lane1- DNA marker

    Techniques Used: Polymerase Chain Reaction, Amplification, Clone Assay, Marker

    30) Product Images from "BioVector, a flexible system for gene specific-expression in plants"

    Article Title: BioVector, a flexible system for gene specific-expression in plants

    Journal: BMC Plant Biology

    doi: 10.1186/1471-2229-13-198

    The general map of GECs. MCS I and MCS II, the multiple cloning sites; att L1 and att L2, recombination sites; ccd B, a negative selection marker for DH5α; CmR, Chloramphenicol resistant in E. coli ; Entr-F and sp6, sequencing primers; the numbers in parenthesis, the position of corresponding restrict enzymes.
    Figure Legend Snippet: The general map of GECs. MCS I and MCS II, the multiple cloning sites; att L1 and att L2, recombination sites; ccd B, a negative selection marker for DH5α; CmR, Chloramphenicol resistant in E. coli ; Entr-F and sp6, sequencing primers; the numbers in parenthesis, the position of corresponding restrict enzymes.

    Techniques Used: Clone Assay, Selection, Marker, Sequencing

    The rationale of BioVector. The LR reaction among GEC, PEC and BDV shows the basic principle of BioVector, in which a gene and a promoter in individual entry clones are positioningly and directionally joined together in a destination vector. att L1/2/3/4, att R1/2/3/4, and att P1/2/3/4, recombination sites; ccd B, a negative selection marker for DH5α; LR Clonase II®, recombination enzyme from Invitrogen.
    Figure Legend Snippet: The rationale of BioVector. The LR reaction among GEC, PEC and BDV shows the basic principle of BioVector, in which a gene and a promoter in individual entry clones are positioningly and directionally joined together in a destination vector. att L1/2/3/4, att R1/2/3/4, and att P1/2/3/4, recombination sites; ccd B, a negative selection marker for DH5α; LR Clonase II®, recombination enzyme from Invitrogen.

    Techniques Used: Clone Assay, Plasmid Preparation, Selection, Marker

    31) Product Images from "Using Colistin as a Trojan Horse: Inactivation of Gram-Negative Bacteria with Chlorophyllin"

    Article Title: Using Colistin as a Trojan Horse: Inactivation of Gram-Negative Bacteria with Chlorophyllin

    Journal: Antibiotics

    doi: 10.3390/antibiotics8040158

    Influence of colistin and chlorophyllin on the growth of Escherichia coli DH5α; ( a ) Growth status of E. coli cultures in lysogeny broth (LB) supplemented with different colistin concentrations after 24 h incubation at 37 °C; ( b ) outer membrane permeability of E. coli in the presence of different colistin concentrations determined by using the NPN uptake assay; ( c , d ) Growth pattern of E. coli in the presence of 10 mg/L chlorophyllin (green line, first column), colistin (yellow lines, central column) and of a combination of both substances (red lines, right column). The black lines describe growth in LB without any supplementation; ( c ) cells were illuminated with a light intensity of 12 mW/cm 2 ; ( d ) cells were incubated in dark. In columns two and three, dotted lines describe growth pattern of cultures in LB + 0.10 µg/mL colistin, dashed lines growth pattern of cultures in LB + 0.25 µg/mL colistin, and solid lines growth pattern of cultures in LB + 0.5 µg/mL colistin. Depicted are measured values (circles) and fitted curves (lines) ± standard deviations (n = 3) showing one representative of three independent experiments. *: p
    Figure Legend Snippet: Influence of colistin and chlorophyllin on the growth of Escherichia coli DH5α; ( a ) Growth status of E. coli cultures in lysogeny broth (LB) supplemented with different colistin concentrations after 24 h incubation at 37 °C; ( b ) outer membrane permeability of E. coli in the presence of different colistin concentrations determined by using the NPN uptake assay; ( c , d ) Growth pattern of E. coli in the presence of 10 mg/L chlorophyllin (green line, first column), colistin (yellow lines, central column) and of a combination of both substances (red lines, right column). The black lines describe growth in LB without any supplementation; ( c ) cells were illuminated with a light intensity of 12 mW/cm 2 ; ( d ) cells were incubated in dark. In columns two and three, dotted lines describe growth pattern of cultures in LB + 0.10 µg/mL colistin, dashed lines growth pattern of cultures in LB + 0.25 µg/mL colistin, and solid lines growth pattern of cultures in LB + 0.5 µg/mL colistin. Depicted are measured values (circles) and fitted curves (lines) ± standard deviations (n = 3) showing one representative of three independent experiments. *: p

    Techniques Used: Incubation, Permeability

    ( a , b ) Broth microdilution plates for MIC determination for colistin/chlorophyllin combinations under ( a ) illuminated and ( b ) dark conditions; ( c ) LB agar plates (n = 3) for the evaluation of CFU ability of wild type and mcr-1 -positive (red arrows) E. coli after incubation to chlorophyllin-colistin combinations. Liquid cultures of E. coli DH5α were supplemented with 10 mg/L chlorophyllin (Chl, samples with green bars) and different colistin concentrations. Cells grew either illuminated with 12 mW/cm 2 or protected from light. Samples (2.5 µL) were drawn at different time points (1 h, 2 h, 3 h, 4 h) and transferred onto LB agar plates. After overnight incubation at 37 °C in the dark, colony growth was analyzed.
    Figure Legend Snippet: ( a , b ) Broth microdilution plates for MIC determination for colistin/chlorophyllin combinations under ( a ) illuminated and ( b ) dark conditions; ( c ) LB agar plates (n = 3) for the evaluation of CFU ability of wild type and mcr-1 -positive (red arrows) E. coli after incubation to chlorophyllin-colistin combinations. Liquid cultures of E. coli DH5α were supplemented with 10 mg/L chlorophyllin (Chl, samples with green bars) and different colistin concentrations. Cells grew either illuminated with 12 mW/cm 2 or protected from light. Samples (2.5 µL) were drawn at different time points (1 h, 2 h, 3 h, 4 h) and transferred onto LB agar plates. After overnight incubation at 37 °C in the dark, colony growth was analyzed.

    Techniques Used: Incubation

    Growth of E. coli DH5α (left column) and mcr-1 -positive E. coli DH5α pGDP2: mcr-1 (right column) supplemented with different colistin concentrations; ( a , b ) cells were illuminated with a light intensity of 12 mW/cm 2 in absence ( a ) and presence ( b ) of 10 mg/L chlorophyllin. The outer membrane permeability was determined by using the NPN uptake assay; ( c , d ) cells were incubated protected from light in absence ( c ) and presence ( d ) of 10 mg/L chlorophyllin. Depicted are measured values ± standard deviations (n = 3) showing one representative of three independent experiments. *: p
    Figure Legend Snippet: Growth of E. coli DH5α (left column) and mcr-1 -positive E. coli DH5α pGDP2: mcr-1 (right column) supplemented with different colistin concentrations; ( a , b ) cells were illuminated with a light intensity of 12 mW/cm 2 in absence ( a ) and presence ( b ) of 10 mg/L chlorophyllin. The outer membrane permeability was determined by using the NPN uptake assay; ( c , d ) cells were incubated protected from light in absence ( c ) and presence ( d ) of 10 mg/L chlorophyllin. Depicted are measured values ± standard deviations (n = 3) showing one representative of three independent experiments. *: p

    Techniques Used: Permeability, Incubation

    32) Product Images from "Decontamination of 16S rRNA gene amplicon sequence datasets based on bacterial load assessment by qPCR"

    Article Title: Decontamination of 16S rRNA gene amplicon sequence datasets based on bacterial load assessment by qPCR

    Journal: BMC Microbiology

    doi: 10.1186/s12866-016-0689-4

    Bacterial load assessed by qPCR. The universal bacterial primers used in qPCR target the V3 segment of the 16S rRNA gene. Bacterial loads were determined using the standard curves obtained with S. aureus MW2 ( a ) or E. coli DH5α ( b ) genomic DNA. The S. aureus and E. coli genomes weigh approximately 2.9 and 4.8 fg and contain six and seven 16S rRNA gene copies, respectively. Each symbol (Exp1, Exp2 and Exp3) corresponds to the series of aliquots processed at a given point and represents the mean of duplicate measurements with relative deviations from the mean
    Figure Legend Snippet: Bacterial load assessed by qPCR. The universal bacterial primers used in qPCR target the V3 segment of the 16S rRNA gene. Bacterial loads were determined using the standard curves obtained with S. aureus MW2 ( a ) or E. coli DH5α ( b ) genomic DNA. The S. aureus and E. coli genomes weigh approximately 2.9 and 4.8 fg and contain six and seven 16S rRNA gene copies, respectively. Each symbol (Exp1, Exp2 and Exp3) corresponds to the series of aliquots processed at a given point and represents the mean of duplicate measurements with relative deviations from the mean

    Techniques Used: Real-time Polymerase Chain Reaction

    Related Articles

    Negative Control:

    Article Title: Enhanced performance of the microalga Chlorella sorokiniana remotely induced by the plant growth-promoting bacteria Azospirillum brasilense and Bacillus pumilus
    Article Snippet: .. In experiments that measured the potential effect of CO2 , Escherichia coli DH5α (Invitrogen, Carlsbad, CA) served as the negative control for microalgal growth and promoting metabolites because it does not have any plant growth-promoting effects; it also served as a positive control in the CO2 experiment because it produces CO2 , as any E. coli . .. For initial culturing of the microalga, 10 mL axenic C. sorokiniana culture, cultivated in sterile mineral medium (C30), was added to a sterile flask containing 90 mL sterile C30 medium, composed of (in g·L−1 ): KNO3 (25), MgSO4 •7H2 O (10), KH2 PO4 (4), K2 HPO4 (1), FeSO4 •7H2 O (1), and (in μg·L−1 ): H3 BO3 (2.86), MnCl2 •4H2 O (1.81), ZnSO4 •7H2 O (0.11), CuSO4 •5H2 O (0.09), NaMoO4 (0.021), pH 5.25 and incubated at 27 ± 2 °C on a rotary shaker at 120 rpm under 60 μmol photon·m−2 ·s−1 continuous light intensity for 6 days .

    Positive Control:

    Article Title: Maturation of molybdoenzymes and its influence on the pathogenesis of non-typeable Haemophilus influenzae
    Article Snippet: .. E. coli Dh5α (Life Technologies) was used as a positive control. ..

    Article Title: Enhanced performance of the microalga Chlorella sorokiniana remotely induced by the plant growth-promoting bacteria Azospirillum brasilense and Bacillus pumilus
    Article Snippet: .. In experiments that measured the potential effect of CO2 , Escherichia coli DH5α (Invitrogen, Carlsbad, CA) served as the negative control for microalgal growth and promoting metabolites because it does not have any plant growth-promoting effects; it also served as a positive control in the CO2 experiment because it produces CO2 , as any E. coli . .. For initial culturing of the microalga, 10 mL axenic C. sorokiniana culture, cultivated in sterile mineral medium (C30), was added to a sterile flask containing 90 mL sterile C30 medium, composed of (in g·L−1 ): KNO3 (25), MgSO4 •7H2 O (10), KH2 PO4 (4), K2 HPO4 (1), FeSO4 •7H2 O (1), and (in μg·L−1 ): H3 BO3 (2.86), MnCl2 •4H2 O (1.81), ZnSO4 •7H2 O (0.11), CuSO4 •5H2 O (0.09), NaMoO4 (0.021), pH 5.25 and incubated at 27 ± 2 °C on a rotary shaker at 120 rpm under 60 μmol photon·m−2 ·s−1 continuous light intensity for 6 days .

    Ligation:

    Article Title: Functions of the Mismatch Repair Gene mutS from Acinetobacter sp. Strain ADP1 †
    Article Snippet: .. Recombinant plasmids were isolated by transforming E. coli DH5α with the appropriate ligation reaction according to the transformation protocol provided by the supplier (Gibco BRL). .. Plasmid pZR7000 was constructed by blunt end ligation of the MUTSF2-MUTSR3 PCR product into the Sma I site of pGEM-3Zf(+).

    Isolation:

    Article Title: Functions of the Mismatch Repair Gene mutS from Acinetobacter sp. Strain ADP1 †
    Article Snippet: .. Recombinant plasmids were isolated by transforming E. coli DH5α with the appropriate ligation reaction according to the transformation protocol provided by the supplier (Gibco BRL). .. Plasmid pZR7000 was constructed by blunt end ligation of the MUTSF2-MUTSR3 PCR product into the Sma I site of pGEM-3Zf(+).

    Construct:

    Article Title: Bacillus thuringiensis subsp. kurstaki HD1 as a factory to synthesize alkali-labile ChiA74?sp chitinase inclusions, Cry crystals and spores for applied use
    Article Snippet: .. All constructs (Figure A) were propagated in E. coli DH5α [supE44, DlacU169 (F80lacZDM15 ) hsdR17 recA1 end A1 gyrA96 thi-1 relA1 ] (Invitrogen, Carlsbad CA, USA) and then used for transforming the acrystalliferous strain of B. thuringiensis subsp. israelensis 4Q7, (hereafter 4Q7), and B. thuringiensis subsp. kurstaki HD1 (hereafter HD1) (Bacillus Genetic Stock Center, Columbus, OH). .. Plasmid pGLO is a vector that harbors the green fluorescent protein (gfp ) gene under the control arabinose (araC ) promoter and contains an ampicillin (bla ) resistance gene marker (Bio-Rad, Hercules CA, USA).

    Article Title: Improving Acetate Tolerance of Escherichia coli by Rewiring Its Global Regulator cAMP Receptor Protein (CRP)
    Article Snippet: .. Materials The host strain E. coli DH5α ∆crp was constructed by knocking out crp from E. coli DH5α (Invitrogen, San Diego, US) according to previously established protocol [ ]. .. Overnight culture was prepared in Luria-Bertani (LB) medium containing 1% tryptone (Oxoid, Hampshire, UK), 0.5% yeast extract (Merck, Damstadt, Germany), and 1% sodium chloride (Merck, Damstadt, Germany).

    Real-time Polymerase Chain Reaction:

    Article Title: Natural microbial communities supporting the transfer of the IncP-1β plasmid pB10 exhibit a higher initial content of plasmids from the same incompatibility group
    Article Snippet: .. Basically, sets of qPCR primers were designed to prime on both sides of a unique junction between building blocks of the element targeted for quantification: a unique assembly between two truncated transposons for plasmid pB10 (target coordinates 45968–46102; GenBank NC_004840), and both sides of a unique 97 kb-deletion in the chromosome of E. coli DH5α (target coordinates on K12 274654–372933; GenBank U00096; ; ). qPCRs were performed in triplicate using a “Step One Plus Real-Time PCR System” (Applied Biosystems, driver: StepOne Software v2.2) with thermocycling conditions set as follows: 2 min at 50°C, then 10 min at 95°C followed by 45 cycles of 15 s at 95°C and 1 min at 60°C. .. Quantifications were carried out from 25 ng of community DNA using a “TaqMan® Universal PCR Master Mix, NoAmpErase® UNG” (Applied Biosystems) as recommended by the manufacturer, with 800 nM of each primer, and 300 nM of TaqMan probe, in a 25 μL reaction volume.

    Expressing:

    Article Title: Amino acid residues 201-205 in C-terminal acidic tail region plays a crucial role in antibacterial activity of HMGB1
    Article Snippet: .. Briefly, the lysates of E. coli DH5α expressing the corresponding proteins were subjected to SDS-PAGE and then transferred onto polyvinylidene difluoride membranes (Pierce). .. After blocked with 5% non-fat milk in phosphate buffered saline containing 0.1% Tween 20 (PBS-T), the membranes were incubated with mouse anti-His monoclonal antibody (Qiagen).

    Transformation Assay:

    Article Title: Requirement of MrpH for Mannose-Resistant Proteus-Like Fimbria-Mediated Hemagglutination by Proteus mirabilis
    Article Snippet: .. The isopropyl-β- d -thiogalactopyranoside (IPTG)-induced cell lysate (about 10 mg protein) of E. coli DH5α transformed with plasmid pMAL-C2 was coupled to an AminoLink Plus column (Pierce Inc., Rockford, Ill.) as specified by the manufacturer. ..

    Article Title: Functions of the Mismatch Repair Gene mutS from Acinetobacter sp. Strain ADP1 †
    Article Snippet: .. Recombinant plasmids were isolated by transforming E. coli DH5α with the appropriate ligation reaction according to the transformation protocol provided by the supplier (Gibco BRL). .. Plasmid pZR7000 was constructed by blunt end ligation of the MUTSF2-MUTSR3 PCR product into the Sma I site of pGEM-3Zf(+).

    Recombinant:

    Article Title: Functions of the Mismatch Repair Gene mutS from Acinetobacter sp. Strain ADP1 †
    Article Snippet: .. Recombinant plasmids were isolated by transforming E. coli DH5α with the appropriate ligation reaction according to the transformation protocol provided by the supplier (Gibco BRL). .. Plasmid pZR7000 was constructed by blunt end ligation of the MUTSF2-MUTSR3 PCR product into the Sma I site of pGEM-3Zf(+).

    SDS Page:

    Article Title: Amino acid residues 201-205 in C-terminal acidic tail region plays a crucial role in antibacterial activity of HMGB1
    Article Snippet: .. Briefly, the lysates of E. coli DH5α expressing the corresponding proteins were subjected to SDS-PAGE and then transferred onto polyvinylidene difluoride membranes (Pierce). .. After blocked with 5% non-fat milk in phosphate buffered saline containing 0.1% Tween 20 (PBS-T), the membranes were incubated with mouse anti-His monoclonal antibody (Qiagen).

    Plasmid Preparation:

    Article Title: Natural microbial communities supporting the transfer of the IncP-1β plasmid pB10 exhibit a higher initial content of plasmids from the same incompatibility group
    Article Snippet: .. Basically, sets of qPCR primers were designed to prime on both sides of a unique junction between building blocks of the element targeted for quantification: a unique assembly between two truncated transposons for plasmid pB10 (target coordinates 45968–46102; GenBank NC_004840), and both sides of a unique 97 kb-deletion in the chromosome of E. coli DH5α (target coordinates on K12 274654–372933; GenBank U00096; ; ). qPCRs were performed in triplicate using a “Step One Plus Real-Time PCR System” (Applied Biosystems, driver: StepOne Software v2.2) with thermocycling conditions set as follows: 2 min at 50°C, then 10 min at 95°C followed by 45 cycles of 15 s at 95°C and 1 min at 60°C. .. Quantifications were carried out from 25 ng of community DNA using a “TaqMan® Universal PCR Master Mix, NoAmpErase® UNG” (Applied Biosystems) as recommended by the manufacturer, with 800 nM of each primer, and 300 nM of TaqMan probe, in a 25 μL reaction volume.

    Article Title: Requirement of MrpH for Mannose-Resistant Proteus-Like Fimbria-Mediated Hemagglutination by Proteus mirabilis
    Article Snippet: .. The isopropyl-β- d -thiogalactopyranoside (IPTG)-induced cell lysate (about 10 mg protein) of E. coli DH5α transformed with plasmid pMAL-C2 was coupled to an AminoLink Plus column (Pierce Inc., Rockford, Ill.) as specified by the manufacturer. ..

    Software:

    Article Title: Natural microbial communities supporting the transfer of the IncP-1β plasmid pB10 exhibit a higher initial content of plasmids from the same incompatibility group
    Article Snippet: .. Basically, sets of qPCR primers were designed to prime on both sides of a unique junction between building blocks of the element targeted for quantification: a unique assembly between two truncated transposons for plasmid pB10 (target coordinates 45968–46102; GenBank NC_004840), and both sides of a unique 97 kb-deletion in the chromosome of E. coli DH5α (target coordinates on K12 274654–372933; GenBank U00096; ; ). qPCRs were performed in triplicate using a “Step One Plus Real-Time PCR System” (Applied Biosystems, driver: StepOne Software v2.2) with thermocycling conditions set as follows: 2 min at 50°C, then 10 min at 95°C followed by 45 cycles of 15 s at 95°C and 1 min at 60°C. .. Quantifications were carried out from 25 ng of community DNA using a “TaqMan® Universal PCR Master Mix, NoAmpErase® UNG” (Applied Biosystems) as recommended by the manufacturer, with 800 nM of each primer, and 300 nM of TaqMan probe, in a 25 μL reaction volume.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Thermo Fisher e coli dh5α
    PCR amplification of E.coli <t>DH5α</t> clones Lane 2 to 7 –positive amplicons Lane1- DNA marker
    E Coli Dh5α, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 170 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli dh5α/product/Thermo Fisher
    Average 94 stars, based on 170 article reviews
    Price from $9.99 to $1999.99
    e coli dh5α - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    93
    Thermo Fisher luciferase expressing e coli dh5α paklux2
    ICE Mh1 PM22 fitness costs, addiction, and consequences for antimicrobial resistance in transconjugants. (A) Growth curves of P. multocida CCUG 17976 (spontaneous rifampin-resistant mutant) and isogenic ICE Mh1 PM22 transconjugant. Mean of 4 biological replicates with SEM. (B) Growth curves of M. haemolytica ATCC 33396 (spontaneous rifampin-resistant mutant) and isogenic ICE Mh1 PM22 transconjugant. Mean of 4 biological replicates with SEM. (C) Luciferase-based competition index from co-cultures of ICE Mh1 PM22 transconjugants and E. coli <t>DH5α</t> harboring the luciferase expression plasmid <t>pAKlux2.</t> Mean of 4 biological replicates with SEM; t-test, ∗ P ≤ 0.05. (D) Long-term repeated passage of ICE Mh1 PM22 transconjugants. Transconjugants were grown in MH (no drug) or MH + 0.5 MIC (subinhibitory; MIC for susceptible P. multocida CCUG 17976 or M. haemolytica ATCC 33396 WT) for 150 days and plated on media with or without selective concentrations of indicated antimicrobials (i.e., 0.5 MIC for non-susceptible isogenic transconjugants). Mean of 3 biological replicates with SEM. (E) Upper panel: growth curves of co-cultures of E. coli DH5α pAKlux2 and E. coli DH5α, P. multocida CCUG 17976 ICE Mh1 PM22 , or M. haemolytica ATCC 33396 ICE Mh1 PM22 in subinhibitory (0.5 MIC for susceptible P. multocida CCUG 17976 or M. haemolytica ATCC 33396) concentrations of oxytetracycline (left) or spectinomycin (right). Mean of 3 biological replicates with SEM. Lower panel: detection of E. coli luciferase production in co-cultures (as above) with either oxytetracycline (left) or spectinomycin (right). (F) Schematic of checkerboard synergy assay for drug interaction screening.
    Luciferase Expressing E Coli Dh5α Paklux2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/luciferase expressing e coli dh5α paklux2/product/Thermo Fisher
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    luciferase expressing e coli dh5α paklux2 - by Bioz Stars, 2020-07
    93/100 stars
      Buy from Supplier

    84
    Thermo Fisher multishot stripwell dh5α t1r competent cells
    Expression of different glycosyl hydrolase (GH) genes from a common expression vector backbone and test of their effects on growth of E. coli <t>DH5α</t> in MOPS minimal medium ( 17 ) and Z. mobilis ZM4 in Zymomonas minimal medium ( 18 ) with glucose or cellobiose as a carbon source. (A) Expression of GH genes in a pVector backbone and a summary of the constructs and their effects on growth in minimal medium supplemented with cellobiose. (B-D) Growth of E. coli DH5α containing plasmids pVector or pCel3A or pGH3 in MOPS minimal medium supplied with 0.4% glucose or cellobiose. (E-G) Growth of Z. mobilis ZM4 containing plasmids pVector or pCel3A or pGH3 in a Zymomonas minimal medium containing 2% glucose or 2% cellobiose. The growth curves are averages of three replicates. *Gene from Cellvibrio japonicus , # Gene from Caulobacter crescentus .
    Multishot Stripwell Dh5α T1r Competent Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/multishot stripwell dh5α t1r competent cells/product/Thermo Fisher
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    multishot stripwell dh5α t1r competent cells - by Bioz Stars, 2020-07
    84/100 stars
      Buy from Supplier

    Image Search Results


    PCR amplification of E.coli DH5α clones Lane 2 to 7 –positive amplicons Lane1- DNA marker

    Journal: Veterinary World

    Article Title: Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a – Escherichia coli DH5α system

    doi: 10.14202/vetworld.2018.557-561

    Figure Lengend Snippet: PCR amplification of E.coli DH5α clones Lane 2 to 7 –positive amplicons Lane1- DNA marker

    Article Snippet: The pET-32a ligA plasmid construct was extracted from the transformed E. coli DH5α using Thermo Scientific Gene JET plasmid Miniprep Kit and subjected to sequencing.

    Techniques: Polymerase Chain Reaction, Amplification, Clone Assay, Marker

    PCR amplification of E.coli DH5α clones Lane 2 to 7 –positive amplicons Lane1- DNA marker

    Journal: Veterinary World

    Article Title: Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a – Escherichia coli DH5α system

    doi: 10.14202/vetworld.2018.557-561

    Figure Lengend Snippet: PCR amplification of E.coli DH5α clones Lane 2 to 7 –positive amplicons Lane1- DNA marker

    Article Snippet: The pET-32a ligA plasmid construct was extracted from the transformed E. coli DH5α using Thermo Scientific Gene JET plasmid Miniprep Kit and subjected to sequencing.

    Techniques: Polymerase Chain Reaction, Amplification, Clone Assay, Marker

    ICE Mh1 PM22 fitness costs, addiction, and consequences for antimicrobial resistance in transconjugants. (A) Growth curves of P. multocida CCUG 17976 (spontaneous rifampin-resistant mutant) and isogenic ICE Mh1 PM22 transconjugant. Mean of 4 biological replicates with SEM. (B) Growth curves of M. haemolytica ATCC 33396 (spontaneous rifampin-resistant mutant) and isogenic ICE Mh1 PM22 transconjugant. Mean of 4 biological replicates with SEM. (C) Luciferase-based competition index from co-cultures of ICE Mh1 PM22 transconjugants and E. coli DH5α harboring the luciferase expression plasmid pAKlux2. Mean of 4 biological replicates with SEM; t-test, ∗ P ≤ 0.05. (D) Long-term repeated passage of ICE Mh1 PM22 transconjugants. Transconjugants were grown in MH (no drug) or MH + 0.5 MIC (subinhibitory; MIC for susceptible P. multocida CCUG 17976 or M. haemolytica ATCC 33396 WT) for 150 days and plated on media with or without selective concentrations of indicated antimicrobials (i.e., 0.5 MIC for non-susceptible isogenic transconjugants). Mean of 3 biological replicates with SEM. (E) Upper panel: growth curves of co-cultures of E. coli DH5α pAKlux2 and E. coli DH5α, P. multocida CCUG 17976 ICE Mh1 PM22 , or M. haemolytica ATCC 33396 ICE Mh1 PM22 in subinhibitory (0.5 MIC for susceptible P. multocida CCUG 17976 or M. haemolytica ATCC 33396) concentrations of oxytetracycline (left) or spectinomycin (right). Mean of 3 biological replicates with SEM. Lower panel: detection of E. coli luciferase production in co-cultures (as above) with either oxytetracycline (left) or spectinomycin (right). (F) Schematic of checkerboard synergy assay for drug interaction screening.

    Journal: Frontiers in Microbiology

    Article Title: Emerging Variants of the Integrative and Conjugant Element ICEMh1 in Livestock Pathogens: Structural Insights, Potential Host Range, and Implications for Bacterial Fitness and Antimicrobial Therapy

    doi: 10.3389/fmicb.2019.02608

    Figure Lengend Snippet: ICE Mh1 PM22 fitness costs, addiction, and consequences for antimicrobial resistance in transconjugants. (A) Growth curves of P. multocida CCUG 17976 (spontaneous rifampin-resistant mutant) and isogenic ICE Mh1 PM22 transconjugant. Mean of 4 biological replicates with SEM. (B) Growth curves of M. haemolytica ATCC 33396 (spontaneous rifampin-resistant mutant) and isogenic ICE Mh1 PM22 transconjugant. Mean of 4 biological replicates with SEM. (C) Luciferase-based competition index from co-cultures of ICE Mh1 PM22 transconjugants and E. coli DH5α harboring the luciferase expression plasmid pAKlux2. Mean of 4 biological replicates with SEM; t-test, ∗ P ≤ 0.05. (D) Long-term repeated passage of ICE Mh1 PM22 transconjugants. Transconjugants were grown in MH (no drug) or MH + 0.5 MIC (subinhibitory; MIC for susceptible P. multocida CCUG 17976 or M. haemolytica ATCC 33396 WT) for 150 days and plated on media with or without selective concentrations of indicated antimicrobials (i.e., 0.5 MIC for non-susceptible isogenic transconjugants). Mean of 3 biological replicates with SEM. (E) Upper panel: growth curves of co-cultures of E. coli DH5α pAKlux2 and E. coli DH5α, P. multocida CCUG 17976 ICE Mh1 PM22 , or M. haemolytica ATCC 33396 ICE Mh1 PM22 in subinhibitory (0.5 MIC for susceptible P. multocida CCUG 17976 or M. haemolytica ATCC 33396) concentrations of oxytetracycline (left) or spectinomycin (right). Mean of 3 biological replicates with SEM. Lower panel: detection of E. coli luciferase production in co-cultures (as above) with either oxytetracycline (left) or spectinomycin (right). (F) Schematic of checkerboard synergy assay for drug interaction screening.

    Article Snippet: For 2-strain co-culture experiments, luciferase-expressing E. coli DH5α pAKlux2 and test strains (including DH5α without pAKlux2 as a control) were each inoculated at an OD600 of 0.0025 (i.e., total OD600 of ∼0.005) into black clear bottom 96-well plates (Nunc, Thermo-Fisher Scientific, Ottawa, ON, Canada).

    Techniques: Mutagenesis, Luciferase, Expressing, Plasmid Preparation

    Expression of different glycosyl hydrolase (GH) genes from a common expression vector backbone and test of their effects on growth of E. coli DH5α in MOPS minimal medium ( 17 ) and Z. mobilis ZM4 in Zymomonas minimal medium ( 18 ) with glucose or cellobiose as a carbon source. (A) Expression of GH genes in a pVector backbone and a summary of the constructs and their effects on growth in minimal medium supplemented with cellobiose. (B-D) Growth of E. coli DH5α containing plasmids pVector or pCel3A or pGH3 in MOPS minimal medium supplied with 0.4% glucose or cellobiose. (E-G) Growth of Z. mobilis ZM4 containing plasmids pVector or pCel3A or pGH3 in a Zymomonas minimal medium containing 2% glucose or 2% cellobiose. The growth curves are averages of three replicates. *Gene from Cellvibrio japonicus , # Gene from Caulobacter crescentus .

    Journal: bioRxiv

    Article Title: Heterologous glycosyl hydrolase expression and cellular reprogramming resembling sucrose-induction enable Zymomonas mobilis growth on cellobiose

    doi: 10.1101/854646

    Figure Lengend Snippet: Expression of different glycosyl hydrolase (GH) genes from a common expression vector backbone and test of their effects on growth of E. coli DH5α in MOPS minimal medium ( 17 ) and Z. mobilis ZM4 in Zymomonas minimal medium ( 18 ) with glucose or cellobiose as a carbon source. (A) Expression of GH genes in a pVector backbone and a summary of the constructs and their effects on growth in minimal medium supplemented with cellobiose. (B-D) Growth of E. coli DH5α containing plasmids pVector or pCel3A or pGH3 in MOPS minimal medium supplied with 0.4% glucose or cellobiose. (E-G) Growth of Z. mobilis ZM4 containing plasmids pVector or pCel3A or pGH3 in a Zymomonas minimal medium containing 2% glucose or 2% cellobiose. The growth curves are averages of three replicates. *Gene from Cellvibrio japonicus , # Gene from Caulobacter crescentus .

    Article Snippet: The E. coli DH10B strain was used for cloning and E. coli DH5α was used for expressing the recombinant plasmids.

    Techniques: Expressing, Plasmid Preparation, Construct