protein expression vector pet15b  (Millipore)


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    Structured Review

    Millipore protein expression vector pet15b
    Protein Expression Vector Pet15b, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/protein expression vector pet15b/product/Millipore
    Average 88 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    protein expression vector pet15b - by Bioz Stars, 2020-04
    88/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae
    Article Snippet: Gste2 was re-amplified from clones of the cDNA extracts using primers that incorporated a 3’Nde I site and a 5’ Bam HI restriction site (Table S1 in ). .. These restriction sites were exploited to clone the Gste2 alleles into protein expression vector pET15b (Novagen) before transformation into E. coli BL21 (DE3) (New England Biolabs).

    Article Title: Two Small (p)ppGpp Synthases in Staphylococcus aureus Mediate Tolerance against Cell Envelope Stress Conditions
    Article Snippet: .. For synthesis of N-terminal His tag fusion proteins of RelP and RelQ, XhoI- and BamHI-digested PCR fragments (for the oligonucleotides used, see Table S2 in the supplemental material) were amplified from S. aureus strain HG001 and cloned into the XhoI-BamHI sites of the protein expression vector pET15b (Novagen), yielding pCG121-RelP and pCG122-RelQ, respectively. .. The verified plasmids were transformed in E. coli strain BL21 (Promega) for further protein expression experiments.

    Article Title: A Glutathione-independent Glyoxalase of the DJ-1 Superfamily Plays an Important Role in Managing Metabolically Generated Methylglyoxal in Candida albicans *
    Article Snippet: .. The PCR product was purified, digested with NdeI and XhoI, and cloned between the corresponding restriction sites of the bacterial protein expression vector pET15b (Novagen). .. The C136S and H137F point mutations were introduced using site-directed mutagenesis, and all constructs were verified using DNA sequencing (Operon).

    Centrifugation:

    Article Title: Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae
    Article Snippet: These restriction sites were exploited to clone the Gste2 alleles into protein expression vector pET15b (Novagen) before transformation into E. coli BL21 (DE3) (New England Biolabs). .. Bacterial lysates were prepared by sonication in buffer TSE (50 mM Tris-HCl pH 7.4, 1 mM EDTA, 150 mM NaCl, 10 mM β-mercaptoethanol (β -ME), 1.25 mM MgCl2 and 250 U benzonase) and cell debris removed through centrifugation (10,000 g for 20 minutes at 4°C) and filtration (0.2 μm filter).

    Amplification:

    Article Title: Two Small (p)ppGpp Synthases in Staphylococcus aureus Mediate Tolerance against Cell Envelope Stress Conditions
    Article Snippet: .. For synthesis of N-terminal His tag fusion proteins of RelP and RelQ, XhoI- and BamHI-digested PCR fragments (for the oligonucleotides used, see Table S2 in the supplemental material) were amplified from S. aureus strain HG001 and cloned into the XhoI-BamHI sites of the protein expression vector pET15b (Novagen), yielding pCG121-RelP and pCG122-RelQ, respectively. .. The verified plasmids were transformed in E. coli strain BL21 (Promega) for further protein expression experiments.

    Filtration:

    Article Title: Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae
    Article Snippet: These restriction sites were exploited to clone the Gste2 alleles into protein expression vector pET15b (Novagen) before transformation into E. coli BL21 (DE3) (New England Biolabs). .. Bacterial lysates were prepared by sonication in buffer TSE (50 mM Tris-HCl pH 7.4, 1 mM EDTA, 150 mM NaCl, 10 mM β-mercaptoethanol (β -ME), 1.25 mM MgCl2 and 250 U benzonase) and cell debris removed through centrifugation (10,000 g for 20 minutes at 4°C) and filtration (0.2 μm filter).

    Mutagenesis:

    Article Title: A Glutathione-independent Glyoxalase of the DJ-1 Superfamily Plays an Important Role in Managing Metabolically Generated Methylglyoxal in Candida albicans *
    Article Snippet: The PCR product was purified, digested with NdeI and XhoI, and cloned between the corresponding restriction sites of the bacterial protein expression vector pET15b (Novagen). .. The C136S and H137F point mutations were introduced using site-directed mutagenesis, and all constructs were verified using DNA sequencing (Operon).

    Construct:

    Article Title: A Glutathione-independent Glyoxalase of the DJ-1 Superfamily Plays an Important Role in Managing Metabolically Generated Methylglyoxal in Candida albicans *
    Article Snippet: The PCR product was purified, digested with NdeI and XhoI, and cloned between the corresponding restriction sites of the bacterial protein expression vector pET15b (Novagen). .. The C136S and H137F point mutations were introduced using site-directed mutagenesis, and all constructs were verified using DNA sequencing (Operon).

    Purification:

    Article Title: Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae
    Article Snippet: These restriction sites were exploited to clone the Gste2 alleles into protein expression vector pET15b (Novagen) before transformation into E. coli BL21 (DE3) (New England Biolabs). .. A pET15b encoded polyhistidine (6XHis) tag was exploited for purification of GSTE2 using nickel affinity chromatography.

    Article Title: Two Small (p)ppGpp Synthases in Staphylococcus aureus Mediate Tolerance against Cell Envelope Stress Conditions
    Article Snippet: Paragraph title: Purification of RelP and RelQ. ... For synthesis of N-terminal His tag fusion proteins of RelP and RelQ, XhoI- and BamHI-digested PCR fragments (for the oligonucleotides used, see Table S2 in the supplemental material) were amplified from S. aureus strain HG001 and cloned into the XhoI-BamHI sites of the protein expression vector pET15b (Novagen), yielding pCG121-RelP and pCG122-RelQ, respectively.

    Article Title: A Glutathione-independent Glyoxalase of the DJ-1 Superfamily Plays an Important Role in Managing Metabolically Generated Methylglyoxal in Candida albicans *
    Article Snippet: .. The PCR product was purified, digested with NdeI and XhoI, and cloned between the corresponding restriction sites of the bacterial protein expression vector pET15b (Novagen). .. The C136S and H137F point mutations were introduced using site-directed mutagenesis, and all constructs were verified using DNA sequencing (Operon).

    Concentration Assay:

    Article Title: Two Small (p)ppGpp Synthases in Staphylococcus aureus Mediate Tolerance against Cell Envelope Stress Conditions
    Article Snippet: For synthesis of N-terminal His tag fusion proteins of RelP and RelQ, XhoI- and BamHI-digested PCR fragments (for the oligonucleotides used, see Table S2 in the supplemental material) were amplified from S. aureus strain HG001 and cloned into the XhoI-BamHI sites of the protein expression vector pET15b (Novagen), yielding pCG121-RelP and pCG122-RelQ, respectively. .. For removal of the imidazole and for concentration, the proteins were ultrafiltrated with ultracentrifugal filter devices (Millipore, Amicon).

    Article Title: A Glutathione-independent Glyoxalase of the DJ-1 Superfamily Plays an Important Role in Managing Metabolically Generated Methylglyoxal in Candida albicans *
    Article Snippet: The PCR product was purified, digested with NdeI and XhoI, and cloned between the corresponding restriction sites of the bacterial protein expression vector pET15b (Novagen). .. Expression of Glx3 was induced by the addition of isopropyl β- d -1-thiogalactopyranoside (Sigma) to a final concentration of 1 m m for 18 h at 25 °C.

    Incubation:

    Article Title: Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae
    Article Snippet: These restriction sites were exploited to clone the Gste2 alleles into protein expression vector pET15b (Novagen) before transformation into E. coli BL21 (DE3) (New England Biolabs). .. Cultures were incubated at 37°C (150RPM) until an optical density of ≈0.8 (wavelength 600 nm) was reached, then protein production was induced by addition of 1 mM isopropy-β-D-thiogalactoside (IPTG) at 30°C (150RPM).

    Affinity Chromatography:

    Article Title: Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae
    Article Snippet: These restriction sites were exploited to clone the Gste2 alleles into protein expression vector pET15b (Novagen) before transformation into E. coli BL21 (DE3) (New England Biolabs). .. A pET15b encoded polyhistidine (6XHis) tag was exploited for purification of GSTE2 using nickel affinity chromatography.

    Activity Assay:

    Article Title: Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae
    Article Snippet: Paragraph title: Recombinant protein expression and DDTase activity screens ... These restriction sites were exploited to clone the Gste2 alleles into protein expression vector pET15b (Novagen) before transformation into E. coli BL21 (DE3) (New England Biolabs).

    Affinity Column:

    Article Title: Two Small (p)ppGpp Synthases in Staphylococcus aureus Mediate Tolerance against Cell Envelope Stress Conditions
    Article Snippet: For synthesis of N-terminal His tag fusion proteins of RelP and RelQ, XhoI- and BamHI-digested PCR fragments (for the oligonucleotides used, see Table S2 in the supplemental material) were amplified from S. aureus strain HG001 and cloned into the XhoI-BamHI sites of the protein expression vector pET15b (Novagen), yielding pCG121-RelP and pCG122-RelQ, respectively. .. His tagged RelP and RelQ fusion proteins were purified on a Ni-nitrilotriacetic acid (NTA) affinity column (GE Healthcare Bio-Sciences Corp.) using imidazole for elution.

    Expressing:

    Article Title: Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae
    Article Snippet: .. These restriction sites were exploited to clone the Gste2 alleles into protein expression vector pET15b (Novagen) before transformation into E. coli BL21 (DE3) (New England Biolabs). .. Cultures were incubated at 37°C (150RPM) until an optical density of ≈0.8 (wavelength 600 nm) was reached, then protein production was induced by addition of 1 mM isopropy-β-D-thiogalactoside (IPTG) at 30°C (150RPM).

    Article Title: Two Small (p)ppGpp Synthases in Staphylococcus aureus Mediate Tolerance against Cell Envelope Stress Conditions
    Article Snippet: .. For synthesis of N-terminal His tag fusion proteins of RelP and RelQ, XhoI- and BamHI-digested PCR fragments (for the oligonucleotides used, see Table S2 in the supplemental material) were amplified from S. aureus strain HG001 and cloned into the XhoI-BamHI sites of the protein expression vector pET15b (Novagen), yielding pCG121-RelP and pCG122-RelQ, respectively. .. The verified plasmids were transformed in E. coli strain BL21 (Promega) for further protein expression experiments.

    Article Title: A Glutathione-independent Glyoxalase of the DJ-1 Superfamily Plays an Important Role in Managing Metabolically Generated Methylglyoxal in Candida albicans *
    Article Snippet: .. The PCR product was purified, digested with NdeI and XhoI, and cloned between the corresponding restriction sites of the bacterial protein expression vector pET15b (Novagen). .. The C136S and H137F point mutations were introduced using site-directed mutagenesis, and all constructs were verified using DNA sequencing (Operon).

    Polymerase Chain Reaction:

    Article Title: Two Small (p)ppGpp Synthases in Staphylococcus aureus Mediate Tolerance against Cell Envelope Stress Conditions
    Article Snippet: .. For synthesis of N-terminal His tag fusion proteins of RelP and RelQ, XhoI- and BamHI-digested PCR fragments (for the oligonucleotides used, see Table S2 in the supplemental material) were amplified from S. aureus strain HG001 and cloned into the XhoI-BamHI sites of the protein expression vector pET15b (Novagen), yielding pCG121-RelP and pCG122-RelQ, respectively. .. The verified plasmids were transformed in E. coli strain BL21 (Promega) for further protein expression experiments.

    Article Title: A Glutathione-independent Glyoxalase of the DJ-1 Superfamily Plays an Important Role in Managing Metabolically Generated Methylglyoxal in Candida albicans *
    Article Snippet: .. The PCR product was purified, digested with NdeI and XhoI, and cloned between the corresponding restriction sites of the bacterial protein expression vector pET15b (Novagen). .. The C136S and H137F point mutations were introduced using site-directed mutagenesis, and all constructs were verified using DNA sequencing (Operon).

    Sonication:

    Article Title: Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae
    Article Snippet: These restriction sites were exploited to clone the Gste2 alleles into protein expression vector pET15b (Novagen) before transformation into E. coli BL21 (DE3) (New England Biolabs). .. Bacterial lysates were prepared by sonication in buffer TSE (50 mM Tris-HCl pH 7.4, 1 mM EDTA, 150 mM NaCl, 10 mM β-mercaptoethanol (β -ME), 1.25 mM MgCl2 and 250 U benzonase) and cell debris removed through centrifugation (10,000 g for 20 minutes at 4°C) and filtration (0.2 μm filter).

    Transformation Assay:

    Article Title: Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae
    Article Snippet: .. These restriction sites were exploited to clone the Gste2 alleles into protein expression vector pET15b (Novagen) before transformation into E. coli BL21 (DE3) (New England Biolabs). .. Cultures were incubated at 37°C (150RPM) until an optical density of ≈0.8 (wavelength 600 nm) was reached, then protein production was induced by addition of 1 mM isopropy-β-D-thiogalactoside (IPTG) at 30°C (150RPM).

    Article Title: Two Small (p)ppGpp Synthases in Staphylococcus aureus Mediate Tolerance against Cell Envelope Stress Conditions
    Article Snippet: For synthesis of N-terminal His tag fusion proteins of RelP and RelQ, XhoI- and BamHI-digested PCR fragments (for the oligonucleotides used, see Table S2 in the supplemental material) were amplified from S. aureus strain HG001 and cloned into the XhoI-BamHI sites of the protein expression vector pET15b (Novagen), yielding pCG121-RelP and pCG122-RelQ, respectively. .. The verified plasmids were transformed in E. coli strain BL21 (Promega) for further protein expression experiments.

    Article Title: A Glutathione-independent Glyoxalase of the DJ-1 Superfamily Plays an Important Role in Managing Metabolically Generated Methylglyoxal in Candida albicans *
    Article Snippet: The PCR product was purified, digested with NdeI and XhoI, and cloned between the corresponding restriction sites of the bacterial protein expression vector pET15b (Novagen). .. E. coli strain BL21 (DE3) (Novagen) was transformed with GLX3 -pET15b and grown at 37 °C in terrific broth medium supplemented with 100 μg/ml ampicillin and with shaking at 270 rpm to an OD600 ∼2.0.

    Binding Assay:

    Article Title: Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae
    Article Snippet: Recombinant protein expression and DDTase activity screens Recombinant protein expression was performed for three Gste2 allelic variants that had non-synonymous changes proximal to the DDT binding site. .. These restriction sites were exploited to clone the Gste2 alleles into protein expression vector pET15b (Novagen) before transformation into E. coli BL21 (DE3) (New England Biolabs).

    Recombinant:

    Article Title: Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae
    Article Snippet: Paragraph title: Recombinant protein expression and DDTase activity screens ... These restriction sites were exploited to clone the Gste2 alleles into protein expression vector pET15b (Novagen) before transformation into E. coli BL21 (DE3) (New England Biolabs).

    Plasmid Preparation:

    Article Title: Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae
    Article Snippet: .. These restriction sites were exploited to clone the Gste2 alleles into protein expression vector pET15b (Novagen) before transformation into E. coli BL21 (DE3) (New England Biolabs). .. Cultures were incubated at 37°C (150RPM) until an optical density of ≈0.8 (wavelength 600 nm) was reached, then protein production was induced by addition of 1 mM isopropy-β-D-thiogalactoside (IPTG) at 30°C (150RPM).

    Article Title: Two Small (p)ppGpp Synthases in Staphylococcus aureus Mediate Tolerance against Cell Envelope Stress Conditions
    Article Snippet: .. For synthesis of N-terminal His tag fusion proteins of RelP and RelQ, XhoI- and BamHI-digested PCR fragments (for the oligonucleotides used, see Table S2 in the supplemental material) were amplified from S. aureus strain HG001 and cloned into the XhoI-BamHI sites of the protein expression vector pET15b (Novagen), yielding pCG121-RelP and pCG122-RelQ, respectively. .. The verified plasmids were transformed in E. coli strain BL21 (Promega) for further protein expression experiments.

    Article Title: A Glutathione-independent Glyoxalase of the DJ-1 Superfamily Plays an Important Role in Managing Metabolically Generated Methylglyoxal in Candida albicans *
    Article Snippet: .. The PCR product was purified, digested with NdeI and XhoI, and cloned between the corresponding restriction sites of the bacterial protein expression vector pET15b (Novagen). .. The C136S and H137F point mutations were introduced using site-directed mutagenesis, and all constructs were verified using DNA sequencing (Operon).

    DNA Sequencing:

    Article Title: A Glutathione-independent Glyoxalase of the DJ-1 Superfamily Plays an Important Role in Managing Metabolically Generated Methylglyoxal in Candida albicans *
    Article Snippet: The PCR product was purified, digested with NdeI and XhoI, and cloned between the corresponding restriction sites of the bacterial protein expression vector pET15b (Novagen). .. The C136S and H137F point mutations were introduced using site-directed mutagenesis, and all constructs were verified using DNA sequencing (Operon).

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  • 87
    Millipore pet15b protein expression vector
    Pet15b Protein Expression Vector, supplied by Millipore, used in various techniques. Bioz Stars score: 87/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pet15b protein expression vector/product/Millipore
    Average 87 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pet15b protein expression vector - by Bioz Stars, 2020-04
    87/100 stars
      Buy from Supplier

    94
    Millipore pet15b expression vector
    Pet15b Expression Vector, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 113 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pet15b expression vector/product/Millipore
    Average 94 stars, based on 113 article reviews
    Price from $9.99 to $1999.99
    pet15b expression vector - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

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