e. coli dh5α Thermo Fisher Search Results


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  • 94
    Thermo Fisher electromax dh5α e electrocompetent e coli
    Electromax Dh5α E Electrocompetent E Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher dh5α e coli
    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.
    Dh5α E Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 495 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher dh5α competent e coli
    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.
    Dh5α Competent E Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher strain dh5α e coli
    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.
    Strain Dh5α E Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Thermo Fisher top10 dh5α competent e coli
    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.
    Top10 Dh5α Competent E Coli, 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
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    96
    Thermo Fisher one shot max efficiency dh5α t1 e coli
    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.
    One Shot Max Efficiency Dh5α T1 E Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 85 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher topo ta cloning kit
    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.
    Topo Ta Cloning Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 53970 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    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.

    Journal: Microbial Cell Factories

    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

    doi: 10.1186/1475-2859-13-15

    Figure Lengend 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.

    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).

    Techniques: 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.

    Journal: Microbial Cell Factories

    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

    doi: 10.1186/1475-2859-13-15

    Figure Lengend 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.

    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).

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

    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.

    Journal: Microbial Cell Factories

    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

    doi: 10.1186/s12934-014-0179-z

    Figure Lengend 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.

    Article Snippet: The plasmid and PCR fragment were ligated using a T4 DNA ligase (New England Biolabs), and transformed to E. coli DH5α cells (Invitrogen) by heat shock.

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