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pet16b  (Millipore)


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    Structured Review

    Millipore pet16b
    Pet16b, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pet16b/pet16b/pmc12205653-248-47-13
    Average 90 stars, based on 1 article reviews
    pet16b - by Bioz Stars, 2026-09
    90/100 stars

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    Related Articles

    Construct:

    Article Title: Neurodevelopmental disorders associated variants in ADAT3 disrupt the activity of the ADAT2/ADAT3 tRNA deaminase complex and impair neuronal migration
    Article Snippet: .. They were further subcloned into psiSTRIKE DCX-IRES-GFP (provided by J. Chelly (IGBMC, Strasbourg, France)) and pnThx (Adat3) , pet16B (Novagen (EMD Millipore)) ( Adat2 ), NeuroD-IRES-GFP ( Adat2 ), pCAGGs-IRES-GFP ( Adat2, Adat3 ) vectors by restriction-ligation. miR1 -Adat3 resistant constructs were obtained by site-directed mutagenesis using the primers indicated in Supplementary Table 3 and subcloned into the psiSTRIKE DCX-IRES-GFP, and pCAGGs-IRES-GFP plasmids. ..

    Article Title: Klebsiella pneumoniae K2 capsular polysaccharide degradation by a bacteriophage depolymerase does not require trimer formation
    Article Snippet: .. The amplified DNA fragment was then inserted into pET16b (Novagen) via Nde I and Hind III cloning sites, and pET21b (Novagen) via Nco I and Xho I cloning sites, resulting in expression constructs that produce the K2-2 with a N-terminal or a C-terminal His-tag, respectively. .. The constructs were then transformed into E. coli expression host BL21(DE3) (Novagen).

    Mutagenesis:

    Article Title: Neurodevelopmental disorders associated variants in ADAT3 disrupt the activity of the ADAT2/ADAT3 tRNA deaminase complex and impair neuronal migration
    Article Snippet: .. They were further subcloned into psiSTRIKE DCX-IRES-GFP (provided by J. Chelly (IGBMC, Strasbourg, France)) and pnThx (Adat3) , pet16B (Novagen (EMD Millipore)) ( Adat2 ), NeuroD-IRES-GFP ( Adat2 ), pCAGGs-IRES-GFP ( Adat2, Adat3 ) vectors by restriction-ligation. miR1 -Adat3 resistant constructs were obtained by site-directed mutagenesis using the primers indicated in Supplementary Table 3 and subcloned into the psiSTRIKE DCX-IRES-GFP, and pCAGGs-IRES-GFP plasmids. ..

    Polymerase Chain Reaction:

    Article Title: Role of the Water-Metal Ion Bridge in Quinolone Interactions with Escherichia coli Gyrase.
    Article Snippet: .. Wild-type GyrA and GyrB genes from E. coli were PCR amplified and cloned into pET16b (Novagen, Madison, WI, USA), which added an N-terminal 10x His-tag. .. GyrA mutants—S83L, D87N, S83F/D87N, and S83A—were generated using a QuikChange Lightning Site-Directed Mutagenesis Kit (Agilent, Santa Clara, CA, USA).

    Article Title: Quantification and structure–function analysis of calpain-1 and calpain-2 protease subunit interactions
    Article Snippet: .. The optimal CAPNS1 PEF(S) biosensor component was PCR amplified and cloned into pET16b (MilliporeSigma; catalog no.: 69662) between NdeI and BamHI; and the optimal CAPN1 or CAPN2 PEF(L) biosensor components were PCR amplified and cloned between BamHI and NdeI, or NdeI and SalI, respectively, in a modified pET16b plasmid (HT29 was a gift from Zongchao Jia, Queen's University) ( ) using the primers shown below. ..

    Amplification:

    Article Title: Role of the Water-Metal Ion Bridge in Quinolone Interactions with Escherichia coli Gyrase.
    Article Snippet: .. Wild-type GyrA and GyrB genes from E. coli were PCR amplified and cloned into pET16b (Novagen, Madison, WI, USA), which added an N-terminal 10x His-tag. .. GyrA mutants—S83L, D87N, S83F/D87N, and S83A—were generated using a QuikChange Lightning Site-Directed Mutagenesis Kit (Agilent, Santa Clara, CA, USA).

    Article Title: Quantification and structure–function analysis of calpain-1 and calpain-2 protease subunit interactions
    Article Snippet: .. The optimal CAPNS1 PEF(S) biosensor component was PCR amplified and cloned into pET16b (MilliporeSigma; catalog no.: 69662) between NdeI and BamHI; and the optimal CAPN1 or CAPN2 PEF(L) biosensor components were PCR amplified and cloned between BamHI and NdeI, or NdeI and SalI, respectively, in a modified pET16b plasmid (HT29 was a gift from Zongchao Jia, Queen's University) ( ) using the primers shown below. ..

    Article Title: Klebsiella pneumoniae K2 capsular polysaccharide degradation by a bacteriophage depolymerase does not require trimer formation
    Article Snippet: .. The amplified DNA fragment was then inserted into pET16b (Novagen) via Nde I and Hind III cloning sites, and pET21b (Novagen) via Nco I and Xho I cloning sites, resulting in expression constructs that produce the K2-2 with a N-terminal or a C-terminal His-tag, respectively. .. The constructs were then transformed into E. coli expression host BL21(DE3) (Novagen).

    Clone Assay:

    Article Title: Role of the Water-Metal Ion Bridge in Quinolone Interactions with Escherichia coli Gyrase.
    Article Snippet: .. Wild-type GyrA and GyrB genes from E. coli were PCR amplified and cloned into pET16b (Novagen, Madison, WI, USA), which added an N-terminal 10x His-tag. .. GyrA mutants—S83L, D87N, S83F/D87N, and S83A—were generated using a QuikChange Lightning Site-Directed Mutagenesis Kit (Agilent, Santa Clara, CA, USA).

    Article Title: Quantification and structure–function analysis of calpain-1 and calpain-2 protease subunit interactions
    Article Snippet: .. The optimal CAPNS1 PEF(S) biosensor component was PCR amplified and cloned into pET16b (MilliporeSigma; catalog no.: 69662) between NdeI and BamHI; and the optimal CAPN1 or CAPN2 PEF(L) biosensor components were PCR amplified and cloned between BamHI and NdeI, or NdeI and SalI, respectively, in a modified pET16b plasmid (HT29 was a gift from Zongchao Jia, Queen's University) ( ) using the primers shown below. ..

    other:

    Article Title: Analysis of Essential Isoprene Metabolic Pathway Proteins in Variovorax sp. Strain WS11
    Article Snippet: Aliquots of the target vector (pET16b [Novagen, UK] for expression of WS11-IsoH, WS11-IsoI, WS11-AldH, AD45-AldH), and pET20MBP (a modified pET20 vector [Novagen, UK], which allows for N-terminal fusion of maltose binding proteins to enhance solubility of heterologous proteins, generously gifted by Nick Burton) for expression of WS11- IsoJ were digested with the appropriate restriction enzymes (Table S1).

    Article Title: A sporulation signature protease is required for assembly of the spore surface layers, germination and host colonization in Clostridioides difficile
    Article Snippet: The resulting fragment was inserted between the BamHI and XhoI sites of pET16b (Novagen) to create pEM6.

    Ubiquitin Proteomics:

    Article Title: Molecular determinants of PQBP1 binding to the HIV-1 capsid lattice.
    Article Snippet: .. Full-length PQBP1 protein was sub-cloned into pET16b (Novagen) with a C-terminal His-tag and Gly-Gly linker. ..

    Modification:

    Article Title: Quantification and structure–function analysis of calpain-1 and calpain-2 protease subunit interactions
    Article Snippet: .. The optimal CAPNS1 PEF(S) biosensor component was PCR amplified and cloned into pET16b (MilliporeSigma; catalog no.: 69662) between NdeI and BamHI; and the optimal CAPN1 or CAPN2 PEF(L) biosensor components were PCR amplified and cloned between BamHI and NdeI, or NdeI and SalI, respectively, in a modified pET16b plasmid (HT29 was a gift from Zongchao Jia, Queen's University) ( ) using the primers shown below. ..

    Plasmid Preparation:

    Article Title: Quantification and structure–function analysis of calpain-1 and calpain-2 protease subunit interactions
    Article Snippet: .. The optimal CAPNS1 PEF(S) biosensor component was PCR amplified and cloned into pET16b (MilliporeSigma; catalog no.: 69662) between NdeI and BamHI; and the optimal CAPN1 or CAPN2 PEF(L) biosensor components were PCR amplified and cloned between BamHI and NdeI, or NdeI and SalI, respectively, in a modified pET16b plasmid (HT29 was a gift from Zongchao Jia, Queen's University) ( ) using the primers shown below. ..

    Cloning:

    Article Title: Klebsiella pneumoniae K2 capsular polysaccharide degradation by a bacteriophage depolymerase does not require trimer formation
    Article Snippet: .. The amplified DNA fragment was then inserted into pET16b (Novagen) via Nde I and Hind III cloning sites, and pET21b (Novagen) via Nco I and Xho I cloning sites, resulting in expression constructs that produce the K2-2 with a N-terminal or a C-terminal His-tag, respectively. .. The constructs were then transformed into E. coli expression host BL21(DE3) (Novagen).

    Expressing:

    Article Title: Klebsiella pneumoniae K2 capsular polysaccharide degradation by a bacteriophage depolymerase does not require trimer formation
    Article Snippet: .. The amplified DNA fragment was then inserted into pET16b (Novagen) via Nde I and Hind III cloning sites, and pET21b (Novagen) via Nco I and Xho I cloning sites, resulting in expression constructs that produce the K2-2 with a N-terminal or a C-terminal His-tag, respectively. .. The constructs were then transformed into E. coli expression host BL21(DE3) (Novagen).



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    Image Search Results


    Fluorescence anisotropy characterizing the protein–DNA interaction between purified Ng Pdc2 DBD and the important cis elements in the NgTHI20 and S cPDC5 promoters. The 55-kDa Ng Pdc2 DBD protein was titrated at different concentrations (µM) with 180 nM of fluorescein-labeled promoter DNA to generate a binding curve and assess the dissociation constant ( K d ) value for the protein–DNA interaction. K d is reported in the figure legend in nanomolars, and the 95% confidence interval is reported in parentheses. As a control, a 38-kDa protein that does not bind DNA was also titrated at different concentrations (µM) with 180 nM of the labeled ScPDC5 promoter DNA. Data shown are the mean and SD of 3 technical replicates.

    Journal: G3: Genes|Genomes|Genetics

    Article Title: Regulation of thiamine and pyruvate decarboxylase genes by Pdc2 in Nakaseomyces glabratus ( Candida glabrata ) is complex

    doi: 10.1093/g3journal/jkae132

    Figure Lengend Snippet: Fluorescence anisotropy characterizing the protein–DNA interaction between purified Ng Pdc2 DBD and the important cis elements in the NgTHI20 and S cPDC5 promoters. The 55-kDa Ng Pdc2 DBD protein was titrated at different concentrations (µM) with 180 nM of fluorescein-labeled promoter DNA to generate a binding curve and assess the dissociation constant ( K d ) value for the protein–DNA interaction. K d is reported in the figure legend in nanomolars, and the 95% confidence interval is reported in parentheses. As a control, a 38-kDa protein that does not bind DNA was also titrated at different concentrations (µM) with 180 nM of the labeled ScPDC5 promoter DNA. Data shown are the mean and SD of 3 technical replicates.

    Article Snippet: To purify the DBD protein using the N-terminal His 10 tag, the Ng Pdc2 DBD-pET16b plasmid was transformed into T7 Express lysY/I q competent Escherichia coli (New England Biolabs).

    Techniques: Fluorescence, Purification, Labeling, Binding Assay, Control

    Pdc2 binding in S. cerevisiae and N. glabratus promoter regions. ChIP was performed on strains where ScPDC2 and NgPDC2 were epitope tagged with c-Myc, as well as on untagged wild-type S. cerevisiae and N. glabratus strains, and grown in high and no thiamine growth conditions. qPCR was used to assess the enrichment of Pdc2 at DNA corresponding to the THI promoters in S. cerevisiae and N. glabratus as well as a gene not regulated by thiamine ( ACT1 ) as a negative control. Data are presented as the ratio of immunoprecipitated DNA from Pdc2-Myc-tagged strains relative to the untagged wild-type strain for each species. Data shown are the average and SE from 3 independent ChIP experiments performed for all strains. A 1-way ANOVA with a post hoc Tukey's multiple comparisons test was performed.

    Journal: G3: Genes|Genomes|Genetics

    Article Title: Regulation of thiamine and pyruvate decarboxylase genes by Pdc2 in Nakaseomyces glabratus ( Candida glabrata ) is complex

    doi: 10.1093/g3journal/jkae132

    Figure Lengend Snippet: Pdc2 binding in S. cerevisiae and N. glabratus promoter regions. ChIP was performed on strains where ScPDC2 and NgPDC2 were epitope tagged with c-Myc, as well as on untagged wild-type S. cerevisiae and N. glabratus strains, and grown in high and no thiamine growth conditions. qPCR was used to assess the enrichment of Pdc2 at DNA corresponding to the THI promoters in S. cerevisiae and N. glabratus as well as a gene not regulated by thiamine ( ACT1 ) as a negative control. Data are presented as the ratio of immunoprecipitated DNA from Pdc2-Myc-tagged strains relative to the untagged wild-type strain for each species. Data shown are the average and SE from 3 independent ChIP experiments performed for all strains. A 1-way ANOVA with a post hoc Tukey's multiple comparisons test was performed.

    Article Snippet: To purify the DBD protein using the N-terminal His 10 tag, the Ng Pdc2 DBD-pET16b plasmid was transformed into T7 Express lysY/I q competent Escherichia coli (New England Biolabs).

    Techniques: Binding Assay, Negative Control, Immunoprecipitation

    Ng Pdc2 is binding to the promoter regions that contain the proposed cis elements for NgTHI20 , NgTHI10 , and NgPET18 , and Ng Thi3 appears to colocalize at these promoters with Pdc2. ChIP was performed on strains where NgPDC2 and NgTHI3 were epitope tagged with c-Myc, as well as on an untagged N. glabratus wild-type strain and grown in no thiamine growth conditions. Enrichment of N. glabratus Pdc2 and Thi3 at promoters was quantified using next-generation sequencing. Coverage for various THI and PDC promoters was plotted against the nucleotide position 1,000-bp upstream and downstream of the ATG, with 6 replicates of N. glabratus wild type and Ng Pdc2-Myc and 3 replicates of Ng Thi3-Myc. There is a peak in coverage for Ng Pdc2-Myc and Ng Thi3-Myc corresponding to the conserved cis sequences found to be required for upregulation of THI20 , THI10 , and PET18 (indicated by a line and asterisk), while the untagged wild-type strain shows no peak. There are no peaks for Ng Pdc2-Myc or Ng Thi3-Myc corresponding to the known cis element important for regulation of the NgPMU3 promoter (indicated by a line and asterisk).

    Journal: G3: Genes|Genomes|Genetics

    Article Title: Regulation of thiamine and pyruvate decarboxylase genes by Pdc2 in Nakaseomyces glabratus ( Candida glabrata ) is complex

    doi: 10.1093/g3journal/jkae132

    Figure Lengend Snippet: Ng Pdc2 is binding to the promoter regions that contain the proposed cis elements for NgTHI20 , NgTHI10 , and NgPET18 , and Ng Thi3 appears to colocalize at these promoters with Pdc2. ChIP was performed on strains where NgPDC2 and NgTHI3 were epitope tagged with c-Myc, as well as on an untagged N. glabratus wild-type strain and grown in no thiamine growth conditions. Enrichment of N. glabratus Pdc2 and Thi3 at promoters was quantified using next-generation sequencing. Coverage for various THI and PDC promoters was plotted against the nucleotide position 1,000-bp upstream and downstream of the ATG, with 6 replicates of N. glabratus wild type and Ng Pdc2-Myc and 3 replicates of Ng Thi3-Myc. There is a peak in coverage for Ng Pdc2-Myc and Ng Thi3-Myc corresponding to the conserved cis sequences found to be required for upregulation of THI20 , THI10 , and PET18 (indicated by a line and asterisk), while the untagged wild-type strain shows no peak. There are no peaks for Ng Pdc2-Myc or Ng Thi3-Myc corresponding to the known cis element important for regulation of the NgPMU3 promoter (indicated by a line and asterisk).

    Article Snippet: To purify the DBD protein using the N-terminal His 10 tag, the Ng Pdc2 DBD-pET16b plasmid was transformed into T7 Express lysY/I q competent Escherichia coli (New England Biolabs).

    Techniques: Binding Assay, Next-Generation Sequencing