bl21 star  (Thermo Fisher)


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    Structured Review

    Thermo Fisher bl21 star
    Immunoblot analysis of protein production using anti-Tdk serum (panels A and C-G) or anti-NilC serum (panel B). E. coli KY895 (panels A, B, F, G), <t>BL21</t> Star™ (DE3) (panel C), KY895 clpP (panel D), or BL21 (DE3) (panel E) carrying plasmids pTara and pET-28a(+) (vector-only; lane 1), pETSTdk or pETSNilC (short-form; lane 2), pETLTdk or pETLNilC (long-form; lane 3), pETMAGPTdk or pETMAGPNilC (D2A change; lane 4), pETMDAPTdk or pETMDAPNilC (G3A change; lane 5) or pETMDGATdk or pETMDGANilC (P4A change; lane 6) was grown overnight in LB/kan/cam and separated cellular proteins were subjected to immunoblot analysis with the indicated antiserum. Arabinose (0.2%) was added to the culture medium for the strains in panels F and G, and IPTG (0.2 mM) was added to the culture medium for the strains in panel G. Similar volumes of cultures were used in each lane but cultures were not normalized for optical density or overall protein concentrations.
    Bl21 Star, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 75 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bl21 star/product/Thermo Fisher
    Average 99 stars, based on 75 article reviews
    Price from $9.99 to $1999.99
    bl21 star - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "An encoded N-terminal extension results in low levels of heterologous protein production in Escherichia coli"

    Article Title: An encoded N-terminal extension results in low levels of heterologous protein production in Escherichia coli

    Journal: Microbial Cell Factories

    doi: 10.1186/1475-2859-4-22

    Immunoblot analysis of protein production using anti-Tdk serum (panels A and C-G) or anti-NilC serum (panel B). E. coli KY895 (panels A, B, F, G), BL21 Star™ (DE3) (panel C), KY895 clpP (panel D), or BL21 (DE3) (panel E) carrying plasmids pTara and pET-28a(+) (vector-only; lane 1), pETSTdk or pETSNilC (short-form; lane 2), pETLTdk or pETLNilC (long-form; lane 3), pETMAGPTdk or pETMAGPNilC (D2A change; lane 4), pETMDAPTdk or pETMDAPNilC (G3A change; lane 5) or pETMDGATdk or pETMDGANilC (P4A change; lane 6) was grown overnight in LB/kan/cam and separated cellular proteins were subjected to immunoblot analysis with the indicated antiserum. Arabinose (0.2%) was added to the culture medium for the strains in panels F and G, and IPTG (0.2 mM) was added to the culture medium for the strains in panel G. Similar volumes of cultures were used in each lane but cultures were not normalized for optical density or overall protein concentrations.
    Figure Legend Snippet: Immunoblot analysis of protein production using anti-Tdk serum (panels A and C-G) or anti-NilC serum (panel B). E. coli KY895 (panels A, B, F, G), BL21 Star™ (DE3) (panel C), KY895 clpP (panel D), or BL21 (DE3) (panel E) carrying plasmids pTara and pET-28a(+) (vector-only; lane 1), pETSTdk or pETSNilC (short-form; lane 2), pETLTdk or pETLNilC (long-form; lane 3), pETMAGPTdk or pETMAGPNilC (D2A change; lane 4), pETMDAPTdk or pETMDAPNilC (G3A change; lane 5) or pETMDGATdk or pETMDGANilC (P4A change; lane 6) was grown overnight in LB/kan/cam and separated cellular proteins were subjected to immunoblot analysis with the indicated antiserum. Arabinose (0.2%) was added to the culture medium for the strains in panels F and G, and IPTG (0.2 mM) was added to the culture medium for the strains in panel G. Similar volumes of cultures were used in each lane but cultures were not normalized for optical density or overall protein concentrations.

    Techniques Used: Positron Emission Tomography, Plasmid Preparation, Chick Chorioallantoic Membrane Assay

    2) Product Images from "An Efficient Strategy for Small-Scale Screening and Production of Archaeal Membrane Transport Proteins in Escherichia coli"

    Article Title: An Efficient Strategy for Small-Scale Screening and Production of Archaeal Membrane Transport Proteins in Escherichia coli

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0076913

    Cell density at induction and harvesting. Expression tests of archaeal transporters were performed in E. coli C43(DE3) and BL21 Star strains. Cells were grown in LB medium, induced with 0.5 mM IPTG at OD 600nm of 0.4-0.8 and harvested 3 hours post-induction. The bars show the OD 600nm at the time of induction (white) and harvesting (black).
    Figure Legend Snippet: Cell density at induction and harvesting. Expression tests of archaeal transporters were performed in E. coli C43(DE3) and BL21 Star strains. Cells were grown in LB medium, induced with 0.5 mM IPTG at OD 600nm of 0.4-0.8 and harvested 3 hours post-induction. The bars show the OD 600nm at the time of induction (white) and harvesting (black).

    Techniques Used: Expressing

    Affinity purification of seven membrane transport proteins. ( A ) Protein purification by Ni-NTA chromatography of 13 constructs expressed in E. coli BL21 Star. SDS-PAGE (top panel) and Western blot (bottom panel) of purified protein. MW - Bio-rad precision plus protein standards. ( B ) Estimated protein yield per litre of cell culture after initial Ni-NTA affinity chromatography. N/D - not determined.
    Figure Legend Snippet: Affinity purification of seven membrane transport proteins. ( A ) Protein purification by Ni-NTA chromatography of 13 constructs expressed in E. coli BL21 Star. SDS-PAGE (top panel) and Western blot (bottom panel) of purified protein. MW - Bio-rad precision plus protein standards. ( B ) Estimated protein yield per litre of cell culture after initial Ni-NTA affinity chromatography. N/D - not determined.

    Techniques Used: Affinity Purification, Protein Purification, Chromatography, Construct, SDS Page, Western Blot, Purification, Cell Culture, Affinity Chromatography

    Western blot analysis of archaeal membrane transport proteins in E. coli membranes. Membrane transport protein genes cloned into pTTQ18, pET52b and pWarf were tested in two E. coli host strains, C43(DE3) and BL21 Star. Membranes were analysed Western blotting. C - C43(DE3), S - BL21 Star, 1 - pTTQ18, 2 - pET52b(+), 3 - pWarf(-).
    Figure Legend Snippet: Western blot analysis of archaeal membrane transport proteins in E. coli membranes. Membrane transport protein genes cloned into pTTQ18, pET52b and pWarf were tested in two E. coli host strains, C43(DE3) and BL21 Star. Membranes were analysed Western blotting. C - C43(DE3), S - BL21 Star, 1 - pTTQ18, 2 - pET52b(+), 3 - pWarf(-).

    Techniques Used: Western Blot, Clone Assay

    SDS-PAGE analysis of archaeal membrane transport proteins in E. coli membranes. Membrane transport protein genes cloned into pTTQ18, pET52b and pWarf were tested in two E. coli host strains, C43(DE3) and BL21 Star. Membranes were analysed by SDS-PAGE. C - C43(DE3), S - BL21 Star, 1 - pTTQ18, 2 - pET52b(+), 3 - pWarf(-). Positions of the His-tagged protein determined by Western blotting (see Figure 3 ) are indicated by: triangle - protein detected by Western blotting but not visible on SDS-PAGE; star - protein detected by Western blotting and visible on SDS-PAGE gels.
    Figure Legend Snippet: SDS-PAGE analysis of archaeal membrane transport proteins in E. coli membranes. Membrane transport protein genes cloned into pTTQ18, pET52b and pWarf were tested in two E. coli host strains, C43(DE3) and BL21 Star. Membranes were analysed by SDS-PAGE. C - C43(DE3), S - BL21 Star, 1 - pTTQ18, 2 - pET52b(+), 3 - pWarf(-). Positions of the His-tagged protein determined by Western blotting (see Figure 3 ) are indicated by: triangle - protein detected by Western blotting but not visible on SDS-PAGE; star - protein detected by Western blotting and visible on SDS-PAGE gels.

    Techniques Used: SDS Page, Clone Assay, Western Blot

    3) Product Images from "Non-pathogenic Escherichia coli biofilms: effects of growth conditions and surface properties on structure and curli gene expression"

    Article Title: Non-pathogenic Escherichia coli biofilms: effects of growth conditions and surface properties on structure and curli gene expression

    Journal: Archives of Microbiology

    doi: 10.1007/s00203-020-01864-5

    Comparison of biofilm formation by three strains (PHL644, BL21 and Nissle 1917) on different substrates. The 6-well plate method was used to compare biofilm formation in M63+ medium on glass, polystyrene (PS), polycarbonate (PC) and stainless steel (SS) coupons. After 3 days, crystal violet staining was used to quantify biofilm formation ( a ). Data shown are the mean ± standard deviation of 3 independent coupons. Significance was determined using a paired two-tailed T -test: * p
    Figure Legend Snippet: Comparison of biofilm formation by three strains (PHL644, BL21 and Nissle 1917) on different substrates. The 6-well plate method was used to compare biofilm formation in M63+ medium on glass, polystyrene (PS), polycarbonate (PC) and stainless steel (SS) coupons. After 3 days, crystal violet staining was used to quantify biofilm formation ( a ). Data shown are the mean ± standard deviation of 3 independent coupons. Significance was determined using a paired two-tailed T -test: * p

    Techniques Used: Staining, Standard Deviation, Two Tailed Test

    4) Product Images from "West Nile alternative open reading frame (N-NS4B/WARF4) is produced in infected West Nile Virus (WNV) cells and induces humoral response in WNV infected individuals"

    Article Title: West Nile alternative open reading frame (N-NS4B/WARF4) is produced in infected West Nile Virus (WNV) cells and induces humoral response in WNV infected individuals

    Journal: Virology Journal

    doi: 10.1186/1743-422X-9-283

    Reactivity of MAb 3A12 with WARF4 recombinant protein. Protein extracts from E. coli BL21 transformed with His-WARF4 and with the empty vector (pRSETC) were analyzed by western blotting. MAb 3A12 reacted with the recombinant His-WARF4 while it did not show reactivity with the crude lysate of E. coli .
    Figure Legend Snippet: Reactivity of MAb 3A12 with WARF4 recombinant protein. Protein extracts from E. coli BL21 transformed with His-WARF4 and with the empty vector (pRSETC) were analyzed by western blotting. MAb 3A12 reacted with the recombinant His-WARF4 while it did not show reactivity with the crude lysate of E. coli .

    Techniques Used: Recombinant, Transformation Assay, Plasmid Preparation, Western Blot

    5) Product Images from "Molecular cloning and characterization of two Helicobacter pylori genes coding for plasminogen-binding proteins"

    Article Title: Molecular cloning and characterization of two Helicobacter pylori genes coding for plasminogen-binding proteins

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.0307329101

    Expression of PgbA and PgbB by H. pylori strains of diverse geographical origins. Shown is a Western blot of H. pylori whole cell lysates using Ab 114, which recognizes both PgbA and -B. Lanes: 1, Poland 43; 2, Turkey 33; 3, Iran 23; 4, Sweden 5; 5, Somalia 3; 6, CCUG 17874; 7, 26695; 8, AH244; 9, E. coli BL21 Star. Molecular mass markers (kDa) are indicated.
    Figure Legend Snippet: Expression of PgbA and PgbB by H. pylori strains of diverse geographical origins. Shown is a Western blot of H. pylori whole cell lysates using Ab 114, which recognizes both PgbA and -B. Lanes: 1, Poland 43; 2, Turkey 33; 3, Iran 23; 4, Sweden 5; 5, Somalia 3; 6, CCUG 17874; 7, 26695; 8, AH244; 9, E. coli BL21 Star. Molecular mass markers (kDa) are indicated.

    Techniques Used: Expressing, Western Blot

    6) Product Images from "Host cell and expression engineering for development of an E. coli ketoreductase catalyst: Enhancement of formate dehydrogenase activity for regeneration of NADH"

    Article Title: Host cell and expression engineering for development of an E. coli ketoreductase catalyst: Enhancement of formate dehydrogenase activity for regeneration of NADH

    Journal: Microbial Cell Factories

    doi: 10.1186/1475-2859-11-7

    Bioreductions of D-xylose catalyzed by whole cells and crude cell extracts of E. coli strains BL21_XR_FDH and Rosetta_XR_2FDH . Type of catalyst is indicated by symbols: ● Rosetta_XR_2FDH crude extract; ○ Rosetta_XR_2FDH whole-cell catalyst; ∇ BL21_XR_FDH crude extract; ▼ BL21_XR_FDH whole-cell catalyst. Reaction conditions: D-xylose (250 mM), sodium formate (300 mM), cells (10 g CDW /L), 30°C.
    Figure Legend Snippet: Bioreductions of D-xylose catalyzed by whole cells and crude cell extracts of E. coli strains BL21_XR_FDH and Rosetta_XR_2FDH . Type of catalyst is indicated by symbols: ● Rosetta_XR_2FDH crude extract; ○ Rosetta_XR_2FDH whole-cell catalyst; ∇ BL21_XR_FDH crude extract; ▼ BL21_XR_FDH whole-cell catalyst. Reaction conditions: D-xylose (250 mM), sodium formate (300 mM), cells (10 g CDW /L), 30°C.

    Techniques Used:

    7) Product Images from "C-terminal domain phosphatase-like family members (AtCPLs) differentially regulate Arabidopsis thaliana abiotic stress signaling, growth, and development"

    Article Title: C-terminal domain phosphatase-like family members (AtCPLs) differentially regulate Arabidopsis thaliana abiotic stress signaling, growth, and development

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.112276199

    AtCPL are phosphatases. AtCPL1 and AtCPL3 cDNAs were placed under the control of the T7 promoter in the vectors pCPL1 and pCPL3 and expressed in E. coli BL21SI and BL21 star cells to produce rCPL1 and rCPL3, respectively. Protein was isolated from cells without and with pCPL1 or pCPL3 plasmid, normalized based on the culture volume (0.25 ml culture per lane), and then fractionated by SDS/PAGE (6.25% gel). Phosphatase activity was visualized by CCD imaging, after impregnating the gel with CDP star substrate (Zymogram). Protein bands were visualized by Coomassie brilliant blue R-25 (CBB) staining after image acquisition. Innate phosphatase activity of bacterial cells is indicated by *. Zymogram indicates the presence of low M r degradation product of rCPL3 in bacterial cells.
    Figure Legend Snippet: AtCPL are phosphatases. AtCPL1 and AtCPL3 cDNAs were placed under the control of the T7 promoter in the vectors pCPL1 and pCPL3 and expressed in E. coli BL21SI and BL21 star cells to produce rCPL1 and rCPL3, respectively. Protein was isolated from cells without and with pCPL1 or pCPL3 plasmid, normalized based on the culture volume (0.25 ml culture per lane), and then fractionated by SDS/PAGE (6.25% gel). Phosphatase activity was visualized by CCD imaging, after impregnating the gel with CDP star substrate (Zymogram). Protein bands were visualized by Coomassie brilliant blue R-25 (CBB) staining after image acquisition. Innate phosphatase activity of bacterial cells is indicated by *. Zymogram indicates the presence of low M r degradation product of rCPL3 in bacterial cells.

    Techniques Used: Isolation, Plasmid Preparation, SDS Page, Activity Assay, Imaging, Staining

    Related Articles

    In Vivo:

    Article Title: Towards characterization of DNA structure under physiological conditions in vivo at the single-molecule level using single-pair FRET
    Article Snippet: .. Incorporation of DNA into E. coli cells The BL21 StarTM(DE3)plysS One Shot (Invitrogen, US) strain of E. coli was used for all in vivo FRET measurements. .. The DNA constructs were introduced into E. coli cells using heat shock with an optimized protocol allowing the introduction of one to five DNA molecules per cell (for details see Supplementary Data ).

    Positron Emission Tomography:

    Article Title: Inhibition of the NKp44-PCNA Immune Checkpoint Using a mAb to PCNA
    Article Snippet: .. Recombinant human PCNA productionThe pET-28 and pMAL-c2x vectors were used to produce soluble human His-tagged and MBP-tagged PCNA respectively in Rosetta™ 2 (DE3) cells. .. Plasmids containing the cDNA or coding sequence of PCNA were transformed into Rosetta™ 2 cells via heat shock, and grown on LB agar plates with kanamycin and chloramphenicol selection.

    Mutagenesis:

    Article Title: Expression, purification and proteomic analysis of recombinant histone H4 acetylated at lysine 16
    Article Snippet: .. Switching to an RNaseE mutant BL21 strain (Invitrogen #C6020-03) substantially improved the expressed protein yield (approximately 200 ug per liter of culture) and allowed its visualization in Coomassie-stained gels ( ). ..

    Construct:

    Article Title: ALS-linked mutations affect UBQLN2 oligomerization and phase separation in a position- and amino acid-dependent manner
    Article Snippet: .. Briefly, the constructs were expressed in Escherichia coli Rosetta 2 (DE3) pLysS cells in Luria-Bertani (LB) broth at 37°C overnight. .. Briefly, NaCl was added to the cleared lysate to the final concentration of 0.5 M-1 M. UBQLN2 droplets were pelleted and then resuspended in 20 mM NaPhosphate, 0.5 mM EDTA (pH 6.8).

    Purification:

    Article Title: Design of a Human Rhinovirus-14 3C Protease-Inducible Caspase-3
    Article Snippet: .. Protein Production and Purification All proteins were produced in BL21(DE3)pLysS E. coli (Thermo Fisher Scientific, Waltham, MA, USA, # C602003). ..

    Produced:

    Article Title: A Two-Step Approach for the Design and Generation of Nanobodies
    Article Snippet: .. Strep-tag-BoNT/A-LC was produced in E. coli BL21(DE3) pLysS (Thermo Fisher Scientific, Waltham, MA, USA, # C602003). ..

    Article Title: Design of a Human Rhinovirus-14 3C Protease-Inducible Caspase-3
    Article Snippet: .. Protein Production and Purification All proteins were produced in BL21(DE3)pLysS E. coli (Thermo Fisher Scientific, Waltham, MA, USA, # C602003). ..

    Transformation Assay:

    Article Title: Allele-specific binding of RNA-binding proteins reveals functional genetic variants in the RNA
    Article Snippet: .. It was transformed into BL21 Star (DE3)-competent cells (Thermo Fisher Scientific, C602003). .. Protein induction was carried out via 1 mM isopropyl β-d -1-thiogalactopyranoside (IPTG) treatment in 50 mL cultured cells (OD = 0.8) for 16 h at 215 rpm at 28 °C.

    Recombinant:

    Article Title: Inhibition of the NKp44-PCNA Immune Checkpoint Using a mAb to PCNA
    Article Snippet: .. Recombinant human PCNA productionThe pET-28 and pMAL-c2x vectors were used to produce soluble human His-tagged and MBP-tagged PCNA respectively in Rosetta™ 2 (DE3) cells. .. Plasmids containing the cDNA or coding sequence of PCNA were transformed into Rosetta™ 2 cells via heat shock, and grown on LB agar plates with kanamycin and chloramphenicol selection.

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  • 99
    Thermo Fisher bl21 cells
    Role of BCAS0292 in epithelial cell attachment. A) Attachment of K56-2 WT, ΔBCAS0292 mutant and Δ0292_0292 complemented strain to CFBE41o − cells at an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain with 20 fields of view counted per data point in three independent experiments, ** p =0.0073. B) Confocal microscopy images representing the attachment of E. coli <t>BL21</t> control and E. coli BL21_BCAS0292 strains to CFBE41o-cells. Bacteria were labelled using a primary anti- E. coli FITC-conjugated antibody (green). Nuclei of CFBE41o-cells were counterstained with DAPI (blue) and actin stained with phalloidin conjugated with Alexa fluor 568. C) Attachment E. coli BL21 control and E. coli BL21_BCAS0292 to CFBE41o-cells, using an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain, with 20 fields of view counted per data point in three independent experiments. D) Confirmation of expression of BCAS0292, following induction in E. coli BL21 cells with 1mM IPTG by Coomassie Blue stained SDS PAGE gel (conditions used for Figure 1C ). Lane 1: MW marker; Lane 2: E. coli BL21 expression control; Lane 3: E. coli BL21_BCAS0292 after incubation with IPTG (1mM) for 2 h.
    Bl21 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bl21 cells/product/Thermo Fisher
    Average 99 stars, based on 28 article reviews
    Price from $9.99 to $1999.99
    bl21 cells - by Bioz Stars, 2020-09
    99/100 stars
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    99
    Thermo Fisher e coli bl21 cells
    Spot assays used for monitoring the growth performance of <t>BL21/pEZYHb</t> and BL21/pEZYHb- ScADH3 transformed E. coli cells on LB plates under cold stress. The growth performance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells on LB plates without supplements or stresses were used as controls. After spotting the sample on LB agar plates, incubation at 4 °C in darkness for 3 days, 7 days, and 14 days was used to study the tolerance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells under cold stress
    E Coli Bl21 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli bl21 cells/product/Thermo Fisher
    Average 99 stars, based on 11 article reviews
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    e coli bl21 cells - by Bioz Stars, 2020-09
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    89
    Thermo Fisher e coli strain bl21 star de3
    Spot assays used for monitoring the growth performance of <t>BL21/pEZYHb</t> and BL21/pEZYHb- ScADH3 transformed E. coli cells on LB plates under cold stress. The growth performance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells on LB plates without supplements or stresses were used as controls. After spotting the sample on LB agar plates, incubation at 4 °C in darkness for 3 days, 7 days, and 14 days was used to study the tolerance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells under cold stress
    E Coli Strain Bl21 Star De3, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli strain bl21 star de3/product/Thermo Fisher
    Average 89 stars, based on 31 article reviews
    Price from $9.99 to $1999.99
    e coli strain bl21 star de3 - by Bioz Stars, 2020-09
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    Image Search Results


    Role of BCAS0292 in epithelial cell attachment. A) Attachment of K56-2 WT, ΔBCAS0292 mutant and Δ0292_0292 complemented strain to CFBE41o − cells at an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain with 20 fields of view counted per data point in three independent experiments, ** p =0.0073. B) Confocal microscopy images representing the attachment of E. coli BL21 control and E. coli BL21_BCAS0292 strains to CFBE41o-cells. Bacteria were labelled using a primary anti- E. coli FITC-conjugated antibody (green). Nuclei of CFBE41o-cells were counterstained with DAPI (blue) and actin stained with phalloidin conjugated with Alexa fluor 568. C) Attachment E. coli BL21 control and E. coli BL21_BCAS0292 to CFBE41o-cells, using an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain, with 20 fields of view counted per data point in three independent experiments. D) Confirmation of expression of BCAS0292, following induction in E. coli BL21 cells with 1mM IPTG by Coomassie Blue stained SDS PAGE gel (conditions used for Figure 1C ). Lane 1: MW marker; Lane 2: E. coli BL21 expression control; Lane 3: E. coli BL21_BCAS0292 after incubation with IPTG (1mM) for 2 h.

    Journal: bioRxiv

    Article Title: The DNA mimic protein BCAS0292 is involved in the regulation of virulence of Burkholderia cenocepacia

    doi: 10.1101/2020.08.06.226779

    Figure Lengend Snippet: Role of BCAS0292 in epithelial cell attachment. A) Attachment of K56-2 WT, ΔBCAS0292 mutant and Δ0292_0292 complemented strain to CFBE41o − cells at an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain with 20 fields of view counted per data point in three independent experiments, ** p =0.0073. B) Confocal microscopy images representing the attachment of E. coli BL21 control and E. coli BL21_BCAS0292 strains to CFBE41o-cells. Bacteria were labelled using a primary anti- E. coli FITC-conjugated antibody (green). Nuclei of CFBE41o-cells were counterstained with DAPI (blue) and actin stained with phalloidin conjugated with Alexa fluor 568. C) Attachment E. coli BL21 control and E. coli BL21_BCAS0292 to CFBE41o-cells, using an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain, with 20 fields of view counted per data point in three independent experiments. D) Confirmation of expression of BCAS0292, following induction in E. coli BL21 cells with 1mM IPTG by Coomassie Blue stained SDS PAGE gel (conditions used for Figure 1C ). Lane 1: MW marker; Lane 2: E. coli BL21 expression control; Lane 3: E. coli BL21_BCAS0292 after incubation with IPTG (1mM) for 2 h.

    Article Snippet: Following the confirmation of recombinant BCAS0292 (rBCAS0292) expression in a pilot study, 1 L cultures of BL21 cells transformed with the expression plasmid, were grown in LB-ampicillin (100 μg/ ml) and induced with IPTG at a final concentration of 1 mM overnight at 25 °C.

    Techniques: Cell Attachment Assay, Mutagenesis, Confocal Microscopy, Staining, Expressing, SDS Page, Marker, Incubation

    Spot assays used for monitoring the growth performance of BL21/pEZYHb and BL21/pEZYHb- ScADH3 transformed E. coli cells on LB plates under cold stress. The growth performance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells on LB plates without supplements or stresses were used as controls. After spotting the sample on LB agar plates, incubation at 4 °C in darkness for 3 days, 7 days, and 14 days was used to study the tolerance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells under cold stress

    Journal: BMC Genomics

    Article Title: The alcohol dehydrogenase gene family in sugarcane and its involvement in cold stress regulation

    doi: 10.1186/s12864-020-06929-9

    Figure Lengend Snippet: Spot assays used for monitoring the growth performance of BL21/pEZYHb and BL21/pEZYHb- ScADH3 transformed E. coli cells on LB plates under cold stress. The growth performance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells on LB plates without supplements or stresses were used as controls. After spotting the sample on LB agar plates, incubation at 4 °C in darkness for 3 days, 7 days, and 14 days was used to study the tolerance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells under cold stress

    Article Snippet: When E. coli BL21 cells containing different vectors were grown to OD600 of 0.6, 1.0 mmol L− 1 isopropyl β-D-thiogalactoside (IPTG) was added to induce protein production.

    Techniques: Transformation Assay, Incubation