e. coli bl21-de3 Search Results


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  • 99
    New England Biolabs e coli bl21
    (A) Extracted ion chromatograms performed on culture supernatant of Escherichia coli <t>BL21</t> <t>DE3</t> pET29a_ agrBD after induction with IPTG (BD+; upper panel) and the synthetic peptide R5T0 (lower panel; R5T0 molecular formula: C 31 H 41 N 5 O 5 S 2 ). (B) Mass spectrometry fragmentation spectra for chromatographic peaks with retention times of 61.2 and 63.9 min (marked with a box in A ). (C,D) Structure and assignment of fragments detected in MS/MS spectra to fragments of R5T0.
    E Coli Bl21, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1143 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher e coli bl21 star de3
    The pGM-5 plasmid in E. coli <t>BL21</t> <t>STAR</t> (DE3) expresses recombinant cCRAMP. (A) Maps of vectors constructed to express (r)cCRAMP as detailed in experimental procedures. (B) E. coli -derived total cell lysates separated in a SDS-PAGE gel containing cCRAMP expressed from pGM-5 (arrow indicates recombinant cCRAMP fusion protein containing a His-tag). (C) A SDS-PAGE gel stained with Coomassie brilliant blue showing purification of cCRAMP from E. coli BL21 STAR (DE3) harboring pGM-5, as detailed in experimental procedures. Protein samples were loaded as follows: Protein bound to the first Ni 2+ -NTA column (lane 2), Ni 2+ -NTA column bound protein digested with thrombin (lane 3), a control protein (Novagen) digested with thrombin (lane 4), thrombin digested protein that did not bind to a second Ni 2+ -NTA column (lane 5), and bound protein from the same second column (lane 6). Arrows show (r)cCRAMP. (D) Further purification of (r)cCRAMP. Thrombin digested protein retained on a 5-kDa size-exclusion column (Lane 2) and (r)CRAMP isolated as a single band after retention on a 3-kDa size-exclusion column (Lane 3). Lane 1 in all gels contains a molecular weight standard.
    E Coli Bl21 Star De3, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 619 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore e coli bl21
    Expression of P. gingivalis thiol peroxidase gene as purified recombinant protein. E. coli <t>BL21/pLysS</t> transformed with the expression vector pET17b containing the thiol peroxidase gene was grown and induced with IPTG. The cells were lysed, and the thiol
    E Coli Bl21, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 10844 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore e coli bl21 de3
    Recombinant AfIlv3A expression, purification and DHAD activity. A. Expression of recombinant Ilv3A protein. E. coli <t>BL21</t> <t>DE3</t> cells were transformed with pET30_Ilv3A, grown until OD600 > 0.5 and then incubated in the presence and absence of 0.5 mM IPTG for 20 h at 20°C. Ilv3A was purified from the bacterial lysate by immobilized metal ion affinity chromatography (IMAC). Protein fractions were separated by polyacrylamide gel electrophoresis on 4–12% Bis-Tris NuPage gels (Invitrogen) followed by staining with GelCode Blue staining reagent (Pierce). The molecular weights in kilo Daltons of marker proteins are indicated. Lane 1, E. coli cell lysate; lane 2, E. coli cell lysate following IPTG induction; lane 3, Ilv3A protein purified by IMAC. B. Dihydroxyacid dehydratase activity of recombinant Ilv3A. 2 µg Ilv3A was assayed in the presence of 0–60 mM L-threonic acid. Initial reaction velocities were calculated and plotted versus substrate concentration.
    E Coli Bl21 De3, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 4332 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher e coli bl21 de3
    Recombinant AfIlv3A expression, purification and DHAD activity. A. Expression of recombinant Ilv3A protein. E. coli <t>BL21</t> <t>DE3</t> cells were transformed with pET30_Ilv3A, grown until OD600 > 0.5 and then incubated in the presence and absence of 0.5 mM IPTG for 20 h at 20°C. Ilv3A was purified from the bacterial lysate by immobilized metal ion affinity chromatography (IMAC). Protein fractions were separated by polyacrylamide gel electrophoresis on 4–12% Bis-Tris NuPage gels (Invitrogen) followed by staining with GelCode Blue staining reagent (Pierce). The molecular weights in kilo Daltons of marker proteins are indicated. Lane 1, E. coli cell lysate; lane 2, E. coli cell lysate following IPTG induction; lane 3, Ilv3A protein purified by IMAC. B. Dihydroxyacid dehydratase activity of recombinant Ilv3A. 2 µg Ilv3A was assayed in the presence of 0–60 mM L-threonic acid. Initial reaction velocities were calculated and plotted versus substrate concentration.
    E Coli Bl21 De3, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 1336 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene e coli bl21 de3
    Recombinant AfIlv3A expression, purification and DHAD activity. A. Expression of recombinant Ilv3A protein. E. coli <t>BL21</t> <t>DE3</t> cells were transformed with pET30_Ilv3A, grown until OD600 > 0.5 and then incubated in the presence and absence of 0.5 mM IPTG for 20 h at 20°C. Ilv3A was purified from the bacterial lysate by immobilized metal ion affinity chromatography (IMAC). Protein fractions were separated by polyacrylamide gel electrophoresis on 4–12% Bis-Tris NuPage gels (Invitrogen) followed by staining with GelCode Blue staining reagent (Pierce). The molecular weights in kilo Daltons of marker proteins are indicated. Lane 1, E. coli cell lysate; lane 2, E. coli cell lysate following IPTG induction; lane 3, Ilv3A protein purified by IMAC. B. Dihydroxyacid dehydratase activity of recombinant Ilv3A. 2 µg Ilv3A was assayed in the presence of 0–60 mM L-threonic acid. Initial reaction velocities were calculated and plotted versus substrate concentration.
    E Coli Bl21 De3, supplied by Stratagene, used in various techniques. Bioz Stars score: 92/100, based on 863 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher one shot bl21 de3 chemically competent e coli
    Recombinant AfIlv3A expression, purification and DHAD activity. A. Expression of recombinant Ilv3A protein. E. coli <t>BL21</t> <t>DE3</t> cells were transformed with pET30_Ilv3A, grown until OD600 > 0.5 and then incubated in the presence and absence of 0.5 mM IPTG for 20 h at 20°C. Ilv3A was purified from the bacterial lysate by immobilized metal ion affinity chromatography (IMAC). Protein fractions were separated by polyacrylamide gel electrophoresis on 4–12% Bis-Tris NuPage gels (Invitrogen) followed by staining with GelCode Blue staining reagent (Pierce). The molecular weights in kilo Daltons of marker proteins are indicated. Lane 1, E. coli cell lysate; lane 2, E. coli cell lysate following IPTG induction; lane 3, Ilv3A protein purified by IMAC. B. Dihydroxyacid dehydratase activity of recombinant Ilv3A. 2 µg Ilv3A was assayed in the presence of 0–60 mM L-threonic acid. Initial reaction velocities were calculated and plotted versus substrate concentration.
    One Shot Bl21 De3 Chemically Competent E Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 194 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher e coli bl21 de3 cells
    Recombinant AfIlv3A expression, purification and DHAD activity. A. Expression of recombinant Ilv3A protein. E. coli <t>BL21</t> <t>DE3</t> cells were transformed with pET30_Ilv3A, grown until OD600 > 0.5 and then incubated in the presence and absence of 0.5 mM IPTG for 20 h at 20°C. Ilv3A was purified from the bacterial lysate by immobilized metal ion affinity chromatography (IMAC). Protein fractions were separated by polyacrylamide gel electrophoresis on 4–12% Bis-Tris NuPage gels (Invitrogen) followed by staining with GelCode Blue staining reagent (Pierce). The molecular weights in kilo Daltons of marker proteins are indicated. Lane 1, E. coli cell lysate; lane 2, E. coli cell lysate following IPTG induction; lane 3, Ilv3A protein purified by IMAC. B. Dihydroxyacid dehydratase activity of recombinant Ilv3A. 2 µg Ilv3A was assayed in the presence of 0–60 mM L-threonic acid. Initial reaction velocities were calculated and plotted versus substrate concentration.
    E Coli Bl21 De3 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1179 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore bl21 de3 chemically competent cells
    Recombinant AfIlv3A expression, purification and DHAD activity. A. Expression of recombinant Ilv3A protein. E. coli <t>BL21</t> <t>DE3</t> cells were transformed with pET30_Ilv3A, grown until OD600 > 0.5 and then incubated in the presence and absence of 0.5 mM IPTG for 20 h at 20°C. Ilv3A was purified from the bacterial lysate by immobilized metal ion affinity chromatography (IMAC). Protein fractions were separated by polyacrylamide gel electrophoresis on 4–12% Bis-Tris NuPage gels (Invitrogen) followed by staining with GelCode Blue staining reagent (Pierce). The molecular weights in kilo Daltons of marker proteins are indicated. Lane 1, E. coli cell lysate; lane 2, E. coli cell lysate following IPTG induction; lane 3, Ilv3A protein purified by IMAC. B. Dihydroxyacid dehydratase activity of recombinant Ilv3A. 2 µg Ilv3A was assayed in the presence of 0–60 mM L-threonic acid. Initial reaction velocities were calculated and plotted versus substrate concentration.
    Bl21 De3 Chemically Competent Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Stratagene e coli bl21 codonplus de3 ril cells
    Overproduction and purification of σ B . (Left panel) SDS-PAGE of protein extracts from E. coli <t>BL21-Codonplus-(DE3)-RIL</t> carrying either pET28-b or pMT01. (Right panel) Immunodetection of σ B with anti-σ B antiserum. Ten micrograms of protein was loaded for each sample. Lane 1, crude protein extract from E. coli carrying pET28-b; lane 2, crude protein extract from E. coli carrying pMT01; lane 3, crude protein extract from E. coli carrying pMT01 after induction with 1 mM IPTG for 2 h; lane 4, inclusion bodies isolated from E. coli carrying pMT01 after induction with 1 mM IPTG for 2 h; lane 5, representative fraction of purified σ B after elution from an Ni 2+ affinity column. The arrow indicates the position of the overproduced σ B protein.
    E Coli Bl21 Codonplus De3 Ril Cells, supplied by Stratagene, used in various techniques. Bioz Stars score: 90/100, based on 632 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare e coli bl21 de3
    Overproduction and purification of σ B . (Left panel) SDS-PAGE of protein extracts from E. coli <t>BL21-Codonplus-(DE3)-RIL</t> carrying either pET28-b or pMT01. (Right panel) Immunodetection of σ B with anti-σ B antiserum. Ten micrograms of protein was loaded for each sample. Lane 1, crude protein extract from E. coli carrying pET28-b; lane 2, crude protein extract from E. coli carrying pMT01; lane 3, crude protein extract from E. coli carrying pMT01 after induction with 1 mM IPTG for 2 h; lane 4, inclusion bodies isolated from E. coli carrying pMT01 after induction with 1 mM IPTG for 2 h; lane 5, representative fraction of purified σ B after elution from an Ni 2+ affinity column. The arrow indicates the position of the overproduced σ B protein.
    E Coli Bl21 De3, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 572 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Stratagene e coli bl21 de3 cells
    Overproduction and purification of σ B . (Left panel) SDS-PAGE of protein extracts from E. coli <t>BL21-Codonplus-(DE3)-RIL</t> carrying either pET28-b or pMT01. (Right panel) Immunodetection of σ B with anti-σ B antiserum. Ten micrograms of protein was loaded for each sample. Lane 1, crude protein extract from E. coli carrying pET28-b; lane 2, crude protein extract from E. coli carrying pMT01; lane 3, crude protein extract from E. coli carrying pMT01 after induction with 1 mM IPTG for 2 h; lane 4, inclusion bodies isolated from E. coli carrying pMT01 after induction with 1 mM IPTG for 2 h; lane 5, representative fraction of purified σ B after elution from an Ni 2+ affinity column. The arrow indicates the position of the overproduced σ B protein.
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    Image Search Results


    (A) Extracted ion chromatograms performed on culture supernatant of Escherichia coli BL21 DE3 pET29a_ agrBD after induction with IPTG (BD+; upper panel) and the synthetic peptide R5T0 (lower panel; R5T0 molecular formula: C 31 H 41 N 5 O 5 S 2 ). (B) Mass spectrometry fragmentation spectra for chromatographic peaks with retention times of 61.2 and 63.9 min (marked with a box in A ). (C,D) Structure and assignment of fragments detected in MS/MS spectra to fragments of R5T0.

    Journal: Frontiers in Microbiology

    Article Title: Identification of the agr Peptide of Listeria monocytogenes

    doi: 10.3389/fmicb.2016.00989

    Figure Lengend Snippet: (A) Extracted ion chromatograms performed on culture supernatant of Escherichia coli BL21 DE3 pET29a_ agrBD after induction with IPTG (BD+; upper panel) and the synthetic peptide R5T0 (lower panel; R5T0 molecular formula: C 31 H 41 N 5 O 5 S 2 ). (B) Mass spectrometry fragmentation spectra for chromatographic peaks with retention times of 61.2 and 63.9 min (marked with a box in A ). (C,D) Structure and assignment of fragments detected in MS/MS spectra to fragments of R5T0.

    Article Snippet: AIP Production in E. coli For heterologous AIP production, pET29a_agrB or pET29a_agrBD were transformed into E. coli BL21(DE3) (New England Biolabs) and transformants were selected on LB agar containing kanamycin.

    Techniques: Mass Spectrometry

    Purification of PilMNO, PilMN, PilM, and PilN. His 6 ‐PilMNO‐Strep (lane 1), His 6 ‐PilMN‐Strep (lane 2), His 6 ‐PilM (lane 3), and PilN‐Strep (lane 4) were purified from Escherichia coli BL21 (DE3) cells as described in Experimental procedures . PilMNO, PilMN, and PilN were isolated from membranes, whereas PilM was isolated from soluble fractions. To separate His 6 ‐PilM (44 kDa), PilN (23 kDa), and PilO‐Strep (22.5 kDa), 4–20% SDS/PAGE was performed (lane 1). All other protein preparations were separated by 14% SDS/PAGE (A). The identity of the proteins was confirmed by western blot using polyclonal antisera against PilM (B) and PilN (C) or using Strep‐Tactin HRP conjugate to detect PilO‐Strep (D). Dotted lines indicate excised lanes from different but comparable gels or blots.

    Journal: FEBS Open Bio

    Article Title: The traffic ATPase PilF interacts with the inner membrane platform of the DNA translocator and type IV pili from Thermus thermophilus

    doi: 10.1002/2211-5463.12548

    Figure Lengend Snippet: Purification of PilMNO, PilMN, PilM, and PilN. His 6 ‐PilMNO‐Strep (lane 1), His 6 ‐PilMN‐Strep (lane 2), His 6 ‐PilM (lane 3), and PilN‐Strep (lane 4) were purified from Escherichia coli BL21 (DE3) cells as described in Experimental procedures . PilMNO, PilMN, and PilN were isolated from membranes, whereas PilM was isolated from soluble fractions. To separate His 6 ‐PilM (44 kDa), PilN (23 kDa), and PilO‐Strep (22.5 kDa), 4–20% SDS/PAGE was performed (lane 1). All other protein preparations were separated by 14% SDS/PAGE (A). The identity of the proteins was confirmed by western blot using polyclonal antisera against PilM (B) and PilN (C) or using Strep‐Tactin HRP conjugate to detect PilO‐Strep (D). Dotted lines indicate excised lanes from different but comparable gels or blots.

    Article Snippet: Organisms and cultivation Escherichia coli DH5α, BL21 (DE3) and T7 Express cells harboring pET28a plasmids (New England Biolabs, Ipswich, MA, USA) were grown at 37 °C in LB medium or 2 × YT medium containing 20 μg·mL−1 kanamycin.

    Techniques: Purification, Isolation, SDS Page, Western Blot

    The pGM-5 plasmid in E. coli BL21 STAR (DE3) expresses recombinant cCRAMP. (A) Maps of vectors constructed to express (r)cCRAMP as detailed in experimental procedures. (B) E. coli -derived total cell lysates separated in a SDS-PAGE gel containing cCRAMP expressed from pGM-5 (arrow indicates recombinant cCRAMP fusion protein containing a His-tag). (C) A SDS-PAGE gel stained with Coomassie brilliant blue showing purification of cCRAMP from E. coli BL21 STAR (DE3) harboring pGM-5, as detailed in experimental procedures. Protein samples were loaded as follows: Protein bound to the first Ni 2+ -NTA column (lane 2), Ni 2+ -NTA column bound protein digested with thrombin (lane 3), a control protein (Novagen) digested with thrombin (lane 4), thrombin digested protein that did not bind to a second Ni 2+ -NTA column (lane 5), and bound protein from the same second column (lane 6). Arrows show (r)cCRAMP. (D) Further purification of (r)cCRAMP. Thrombin digested protein retained on a 5-kDa size-exclusion column (Lane 2) and (r)CRAMP isolated as a single band after retention on a 3-kDa size-exclusion column (Lane 3). Lane 1 in all gels contains a molecular weight standard.

    Journal: Molecular immunology

    Article Title: A member of the cathelicidin family of antimicrobial peptides is produced in the upper airway of the chinchilla and its mRNA expression is altered by common viral and bacterial co-pathogens of otitis media

    doi: 10.1016/j.molimm.2006.10.008

    Figure Lengend Snippet: The pGM-5 plasmid in E. coli BL21 STAR (DE3) expresses recombinant cCRAMP. (A) Maps of vectors constructed to express (r)cCRAMP as detailed in experimental procedures. (B) E. coli -derived total cell lysates separated in a SDS-PAGE gel containing cCRAMP expressed from pGM-5 (arrow indicates recombinant cCRAMP fusion protein containing a His-tag). (C) A SDS-PAGE gel stained with Coomassie brilliant blue showing purification of cCRAMP from E. coli BL21 STAR (DE3) harboring pGM-5, as detailed in experimental procedures. Protein samples were loaded as follows: Protein bound to the first Ni 2+ -NTA column (lane 2), Ni 2+ -NTA column bound protein digested with thrombin (lane 3), a control protein (Novagen) digested with thrombin (lane 4), thrombin digested protein that did not bind to a second Ni 2+ -NTA column (lane 5), and bound protein from the same second column (lane 6). Arrows show (r)cCRAMP. (D) Further purification of (r)cCRAMP. Thrombin digested protein retained on a 5-kDa size-exclusion column (Lane 2) and (r)CRAMP isolated as a single band after retention on a 3-kDa size-exclusion column (Lane 3). Lane 1 in all gels contains a molecular weight standard.

    Article Snippet: This DNA construct was transformed into E. coli BL21 trxB (DE3) (Novagen), E. coli Rosetta (DE3) (Novagen) and E. coli BL21 STAR (DE3) (Invitrogen), and the appearance of an IPTG-inducible protein band of appropriate molecular mass was evaluated in Coomassie brilliant blue-stained SDS-PAGE gels of total cell lysates.

    Techniques: Plasmid Preparation, Recombinant, Construct, Derivative Assay, SDS Page, Staining, Purification, Isolation, Molecular Weight

    Expression of P. gingivalis thiol peroxidase gene as purified recombinant protein. E. coli BL21/pLysS transformed with the expression vector pET17b containing the thiol peroxidase gene was grown and induced with IPTG. The cells were lysed, and the thiol

    Journal:

    Article Title: T-Cell Expression Cloning of Porphyromonas gingivalis Genes Coding for T Helper-Biased Immune Responses during Infection

    doi: 10.1128/IAI.02029-05

    Figure Lengend Snippet: Expression of P. gingivalis thiol peroxidase gene as purified recombinant protein. E. coli BL21/pLysS transformed with the expression vector pET17b containing the thiol peroxidase gene was grown and induced with IPTG. The cells were lysed, and the thiol

    Article Snippet: Ligation products were subsequently transformed into E. coli BL21(DE3)/pLysS host cells (Novagen) for expression.

    Techniques: Expressing, Purification, Recombinant, Transformation Assay, Plasmid Preparation

    Recombinant AfIlv3A expression, purification and DHAD activity. A. Expression of recombinant Ilv3A protein. E. coli BL21 DE3 cells were transformed with pET30_Ilv3A, grown until OD600 > 0.5 and then incubated in the presence and absence of 0.5 mM IPTG for 20 h at 20°C. Ilv3A was purified from the bacterial lysate by immobilized metal ion affinity chromatography (IMAC). Protein fractions were separated by polyacrylamide gel electrophoresis on 4–12% Bis-Tris NuPage gels (Invitrogen) followed by staining with GelCode Blue staining reagent (Pierce). The molecular weights in kilo Daltons of marker proteins are indicated. Lane 1, E. coli cell lysate; lane 2, E. coli cell lysate following IPTG induction; lane 3, Ilv3A protein purified by IMAC. B. Dihydroxyacid dehydratase activity of recombinant Ilv3A. 2 µg Ilv3A was assayed in the presence of 0–60 mM L-threonic acid. Initial reaction velocities were calculated and plotted versus substrate concentration.

    Journal: PLoS ONE

    Article Title: The Aspergillus fumigatus Dihydroxyacid Dehydratase Ilv3A/IlvC Is Required for Full Virulence

    doi: 10.1371/journal.pone.0043559

    Figure Lengend Snippet: Recombinant AfIlv3A expression, purification and DHAD activity. A. Expression of recombinant Ilv3A protein. E. coli BL21 DE3 cells were transformed with pET30_Ilv3A, grown until OD600 > 0.5 and then incubated in the presence and absence of 0.5 mM IPTG for 20 h at 20°C. Ilv3A was purified from the bacterial lysate by immobilized metal ion affinity chromatography (IMAC). Protein fractions were separated by polyacrylamide gel electrophoresis on 4–12% Bis-Tris NuPage gels (Invitrogen) followed by staining with GelCode Blue staining reagent (Pierce). The molecular weights in kilo Daltons of marker proteins are indicated. Lane 1, E. coli cell lysate; lane 2, E. coli cell lysate following IPTG induction; lane 3, Ilv3A protein purified by IMAC. B. Dihydroxyacid dehydratase activity of recombinant Ilv3A. 2 µg Ilv3A was assayed in the presence of 0–60 mM L-threonic acid. Initial reaction velocities were calculated and plotted versus substrate concentration.

    Article Snippet: Sequencing confirmed that the resulting vector, pET30_Ilv3A, was correctly constructed. pET30_Ilv3A was transformed into E. coli BL21 DE3 (Novagen) and expression was induced by IPTG as directed in the manufacturers protocol.

    Techniques: Recombinant, Expressing, Purification, Activity Assay, Transformation Assay, Incubation, Affinity Chromatography, Polyacrylamide Gel Electrophoresis, Staining, Marker, Concentration Assay

    Electrophoretic profile of total cellular proteins obtained from E. coli DE3 strains BL21, Rosetta-2, C41, and C43 (labeled above each lane) expressing TM4-Cx43CT after IPTG induction for 4 hrs (panel A, lanes 3, 5, 7, and 9). Lanes 2, 4, 6 and 8 are controls (no IPTG). Lane 1 contains the protein molecular weight marker. B) The presence of the TM4-Cx43CT polypeptide was confirmed by western blot analysis using a Zymed Laboratories Inc. polyclonal anti-Cx43CT antibody. The blots correspond to lanes 6 and 7 from panel A. The arrows represent the location of the TM4-Cx43CT monomer. The asterisk represents TM4-Cx43CT dimers.

    Journal: Protein expression and purification

    Article Title: Purification and Reconstitution of the Connexin43 Carboxyl Terminus attached to the 4th Transmembrane Domain in Detergent Micelles *

    doi: 10.1016/j.pep.2008.01.023

    Figure Lengend Snippet: Electrophoretic profile of total cellular proteins obtained from E. coli DE3 strains BL21, Rosetta-2, C41, and C43 (labeled above each lane) expressing TM4-Cx43CT after IPTG induction for 4 hrs (panel A, lanes 3, 5, 7, and 9). Lanes 2, 4, 6 and 8 are controls (no IPTG). Lane 1 contains the protein molecular weight marker. B) The presence of the TM4-Cx43CT polypeptide was confirmed by western blot analysis using a Zymed Laboratories Inc. polyclonal anti-Cx43CT antibody. The blots correspond to lanes 6 and 7 from panel A. The arrows represent the location of the TM4-Cx43CT monomer. The asterisk represents TM4-Cx43CT dimers.

    Article Snippet: The E. coli strains BL21(DE3) (Novagen), Rosetta-2(DE3) (Novagen), C41(DE3) (Lucigen, ( )), and C43(DE3) (Lucigen, ( )) were transformed with the TM4-Cx43CT expression plasmid (see above) and then inoculated into 1 L of Luria Bertani medium (LB), enriched minimal media , or ISOGRO media (Sigma-Aldrich).

    Techniques: Labeling, Expressing, Molecular Weight, Marker, Western Blot

    Overexpression of the recombinant T. pallidum Gpd and analysis of anti-Gpd immunoreactivity. (A) Coomassie blue-stained SDS-PAGE analysis of E. coli BL21 (DE3) pLysS expressing either the Gpd-pET-3a construct (lane 1, crude lysate; lane 2, soluble fraction; lane 3, insoluble fraction) or the pET-3a vector alone (lane 4, insoluble fraction). (B) Immunoblot analysis of anti-Gpd immunoreactivity on purified inclusion bodies from E. coli BL21 (DE3) pLysS expressing the Gpd-pET-3a construct. Lanes: 1, anti-Gpd polyclonal antiserum, 2, E. coli -adsorbed anti-Gpd polyclonal antiserum. Each lane contains approximately 2 μg of total bacterial lysate and soluble or insoluble bacterial fractions, and molecular mass standards in kilodaltons are indicated at the left of each panel. In each panel, the recombinant T. pallidum Gpd is indicated by an arrow.

    Journal: Infection and Immunity

    Article Title: Function and Protective Capacity of Treponema pallidum subsp. pallidum Glycerophosphodiester Phosphodiesterase

    doi:

    Figure Lengend Snippet: Overexpression of the recombinant T. pallidum Gpd and analysis of anti-Gpd immunoreactivity. (A) Coomassie blue-stained SDS-PAGE analysis of E. coli BL21 (DE3) pLysS expressing either the Gpd-pET-3a construct (lane 1, crude lysate; lane 2, soluble fraction; lane 3, insoluble fraction) or the pET-3a vector alone (lane 4, insoluble fraction). (B) Immunoblot analysis of anti-Gpd immunoreactivity on purified inclusion bodies from E. coli BL21 (DE3) pLysS expressing the Gpd-pET-3a construct. Lanes: 1, anti-Gpd polyclonal antiserum, 2, E. coli -adsorbed anti-Gpd polyclonal antiserum. Each lane contains approximately 2 μg of total bacterial lysate and soluble or insoluble bacterial fractions, and molecular mass standards in kilodaltons are indicated at the left of each panel. In each panel, the recombinant T. pallidum Gpd is indicated by an arrow.

    Article Snippet: Anti-Gpd polyclonal antiserum was raised in a New Zealand White rabbit by immunizing three times with 100 μg of each of the inclusion bodies purified from E. coli BL21 (DE3) pLysS transformed with the Gpd-pET-3a construct, emulsified in the Ribi adjuvant MPL + TDM + CWS (monophosphoryl lipid A + trehalose dicorynomycolate + cell wall skeleton) (Sigma).

    Techniques: Over Expression, Recombinant, Staining, SDS Page, Expressing, Positron Emission Tomography, Construct, Plasmid Preparation, Purification

    Immunoblot analyses of the Ig-binding capability of recombinant T. pallidum Gpd. Shown are inclusion bodies purified from E. coli BL21 (DE3) pLysS transformed with either the pET-3a-Gpd construct (lanes 1, 3, 5, and 7) or with the pET-3a vector alone (lanes 2, 4, 6, and 8). Molecular mass standards in kilodaltons are indicated on the left of each panel. (A) Specificity of recombinant T. pallidum Gpd for binding various Ig classes. Lanes: 1 and 2, human IgA and goat F(ab′) 2 anti-human IgA (alpha chain specific); 3 and 4, human IgD and goat F(ab′) 2 anti-human IgD (delta chain specific); 5 and 6, human IgG and goat F(ab′) 2 anti-human IgG (gamma chain specific); 7 and 8, human IgM and goat F(ab′) 2 anti-human IgM (mu chain specific). (B) Investigation of the binding specificity of recombinant T. pallidum Gpd for human IgG. In all cases the secondary antibody used was goat F(ab′) 2 anti-human IgG (gamma chain specific). Lanes: 1 and 2, intact human IgG; 3 and 4, human IgG Fc fragment; 5 and 6, human IgG Fab fragment; 7 and 8, no primary Ig.

    Journal: Infection and Immunity

    Article Title: Function and Protective Capacity of Treponema pallidum subsp. pallidum Glycerophosphodiester Phosphodiesterase

    doi:

    Figure Lengend Snippet: Immunoblot analyses of the Ig-binding capability of recombinant T. pallidum Gpd. Shown are inclusion bodies purified from E. coli BL21 (DE3) pLysS transformed with either the pET-3a-Gpd construct (lanes 1, 3, 5, and 7) or with the pET-3a vector alone (lanes 2, 4, 6, and 8). Molecular mass standards in kilodaltons are indicated on the left of each panel. (A) Specificity of recombinant T. pallidum Gpd for binding various Ig classes. Lanes: 1 and 2, human IgA and goat F(ab′) 2 anti-human IgA (alpha chain specific); 3 and 4, human IgD and goat F(ab′) 2 anti-human IgD (delta chain specific); 5 and 6, human IgG and goat F(ab′) 2 anti-human IgG (gamma chain specific); 7 and 8, human IgM and goat F(ab′) 2 anti-human IgM (mu chain specific). (B) Investigation of the binding specificity of recombinant T. pallidum Gpd for human IgG. In all cases the secondary antibody used was goat F(ab′) 2 anti-human IgG (gamma chain specific). Lanes: 1 and 2, intact human IgG; 3 and 4, human IgG Fc fragment; 5 and 6, human IgG Fab fragment; 7 and 8, no primary Ig.

    Article Snippet: Anti-Gpd polyclonal antiserum was raised in a New Zealand White rabbit by immunizing three times with 100 μg of each of the inclusion bodies purified from E. coli BL21 (DE3) pLysS transformed with the Gpd-pET-3a construct, emulsified in the Ribi adjuvant MPL + TDM + CWS (monophosphoryl lipid A + trehalose dicorynomycolate + cell wall skeleton) (Sigma).

    Techniques: Binding Assay, Recombinant, Purification, Transformation Assay, Positron Emission Tomography, Construct, Plasmid Preparation

    Overproduction and purification of σ B . (Left panel) SDS-PAGE of protein extracts from E. coli BL21-Codonplus-(DE3)-RIL carrying either pET28-b or pMT01. (Right panel) Immunodetection of σ B with anti-σ B antiserum. Ten micrograms of protein was loaded for each sample. Lane 1, crude protein extract from E. coli carrying pET28-b; lane 2, crude protein extract from E. coli carrying pMT01; lane 3, crude protein extract from E. coli carrying pMT01 after induction with 1 mM IPTG for 2 h; lane 4, inclusion bodies isolated from E. coli carrying pMT01 after induction with 1 mM IPTG for 2 h; lane 5, representative fraction of purified σ B after elution from an Ni 2+ affinity column. The arrow indicates the position of the overproduced σ B protein.

    Journal: Journal of Bacteriology

    Article Title: The Alternative Sigma Factor ?B of Bacillus cereus: Response to Stress and Role in Heat Adaptation

    doi: 10.1128/JB.186.2.316-325.2004

    Figure Lengend Snippet: Overproduction and purification of σ B . (Left panel) SDS-PAGE of protein extracts from E. coli BL21-Codonplus-(DE3)-RIL carrying either pET28-b or pMT01. (Right panel) Immunodetection of σ B with anti-σ B antiserum. Ten micrograms of protein was loaded for each sample. Lane 1, crude protein extract from E. coli carrying pET28-b; lane 2, crude protein extract from E. coli carrying pMT01; lane 3, crude protein extract from E. coli carrying pMT01 after induction with 1 mM IPTG for 2 h; lane 4, inclusion bodies isolated from E. coli carrying pMT01 after induction with 1 mM IPTG for 2 h; lane 5, representative fraction of purified σ B after elution from an Ni 2+ affinity column. The arrow indicates the position of the overproduced σ B protein.

    Article Snippet: E. coli BL21-Codonplus-(DE3)-RIL (Stratagene, La Jolla, Calif.) was used as the host for SigB overproduction.

    Techniques: Purification, SDS Page, Immunodetection, Isolation, Affinity Column