pbr322 vector  (New England Biolabs)


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    Name:
    pBR322 Vector
    Description:
    pBR322 Vector 250 ug
    Catalog Number:
    n3033l
    Price:
    302
    Size:
    250 ug
    Category:
    Vectors Plasmids
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    Structured Review

    New England Biolabs pbr322 vector
    pBR322 Vector
    pBR322 Vector 250 ug
    https://www.bioz.com/result/pbr322 vector/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pbr322 vector - by Bioz Stars, 2020-07
    95/100 stars

    Images

    1) Product Images from "Investigation into the antimicrobial action and mechanism of a novel endogenous peptide β-casein 197 from human milk"

    Article Title: Investigation into the antimicrobial action and mechanism of a novel endogenous peptide β-casein 197 from human milk

    Journal: AMB Express

    doi: 10.1186/s13568-017-0409-y

    The DNA-binding properties of β-Casein 197. a ProtParam analysis of predicted binding residues of β-casein 197, predicted binding residues was labeled with a red “+”. b Gel retardation assays. Binding was assayed by the migration of pDNA. Various concentrations of peptides were incubated with 5 µL of 25 µg mL −1 pBR322 vector from E. coli at 37 °C for 1 h, and then the reaction mixtures were applied to 0.7% agarose gel electrophoresis. Lane M DNA marker DL 10,000; Lane 1 5 µL pDNA as control; Lanes 2–6 mixture of pDNA and various concentrations of peptides (12.5, 25, 50, 100 and 500 µg mL −1 )
    Figure Legend Snippet: The DNA-binding properties of β-Casein 197. a ProtParam analysis of predicted binding residues of β-casein 197, predicted binding residues was labeled with a red “+”. b Gel retardation assays. Binding was assayed by the migration of pDNA. Various concentrations of peptides were incubated with 5 µL of 25 µg mL −1 pBR322 vector from E. coli at 37 °C for 1 h, and then the reaction mixtures were applied to 0.7% agarose gel electrophoresis. Lane M DNA marker DL 10,000; Lane 1 5 µL pDNA as control; Lanes 2–6 mixture of pDNA and various concentrations of peptides (12.5, 25, 50, 100 and 500 µg mL −1 )

    Techniques Used: Binding Assay, Labeling, Electrophoretic Mobility Shift Assay, Migration, Incubation, Plasmid Preparation, Agarose Gel Electrophoresis, Marker

    2) Product Images from "Dataset on the effects of spermidine on linking number differences between histone H1-free and histone H1-bound circular polynucleosomes"

    Article Title: Dataset on the effects of spermidine on linking number differences between histone H1-free and histone H1-bound circular polynucleosomes

    Journal: Data in Brief

    doi: 10.1016/j.dib.2018.01.091

    (a) Illustration of our preparations of Structure 26 and Structure 28 in the presence of 5 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 26 and Structure 28. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 26 and Lane 4: Structure 28. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 5 c, which gave rise to − 5.20 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 5 e, which gave rise to − 5.32 as its mean value of ΔLk .
    Figure Legend Snippet: (a) Illustration of our preparations of Structure 26 and Structure 28 in the presence of 5 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 26 and Structure 28. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 26 and Lane 4: Structure 28. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 5 c, which gave rise to − 5.20 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 5 e, which gave rise to − 5.32 as its mean value of ΔLk .

    Techniques Used: Agarose Gel Electrophoresis, Molecular Weight

    (a) Illustration of our preparations of Structure 10 and Structure 12 in the presence of 0.75 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 10 and Structure 12. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 10 and Lane 4: Structure 12. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 1 c, which gave rise to − 6.34 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 1 e, which gave rise to − 6.07 as its mean value of ΔLk .
    Figure Legend Snippet: (a) Illustration of our preparations of Structure 10 and Structure 12 in the presence of 0.75 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 10 and Structure 12. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 10 and Lane 4: Structure 12. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 1 c, which gave rise to − 6.34 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 1 e, which gave rise to − 6.07 as its mean value of ΔLk .

    Techniques Used: Agarose Gel Electrophoresis, Molecular Weight

    (a) Illustration of our preparations of Structure 18 and Structure 20 in the presence of 2 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 18 and Structure 20. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 18 and Lane 4: Structure 20. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 3 c, which gave rise to − 6.41 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 3 e, which gave rise to − 5.98 as its mean value of ΔLk .
    Figure Legend Snippet: (a) Illustration of our preparations of Structure 18 and Structure 20 in the presence of 2 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 18 and Structure 20. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 18 and Lane 4: Structure 20. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 3 c, which gave rise to − 6.41 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 3 e, which gave rise to − 5.98 as its mean value of ΔLk .

    Techniques Used: Agarose Gel Electrophoresis, Molecular Weight

    (a) Illustration of our preparations of Structure 14 and Structure 16 in the presence of 0 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 14 and Structure 16. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 14 and Lane 4: Structure 16. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 2 c, which gave rise to − 6.01 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 2 e, which gave rise to − 5.98 as its mean value of ΔLk .
    Figure Legend Snippet: (a) Illustration of our preparations of Structure 14 and Structure 16 in the presence of 0 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 14 and Structure 16. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 14 and Lane 4: Structure 16. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 2 c, which gave rise to − 6.01 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 2 e, which gave rise to − 5.98 as its mean value of ΔLk .

    Techniques Used: Agarose Gel Electrophoresis, Molecular Weight

    (a) Illustration of our preparations of Structure 22 and Structure 24 in the presence of 3 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 22 and Structure 24. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 22 and Lane 4: Structure 24. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 4 c, which gave rise to − 5.48 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 4 e, which gave rise to − 5.19 as its mean value of ΔLk .
    Figure Legend Snippet: (a) Illustration of our preparations of Structure 22 and Structure 24 in the presence of 3 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 22 and Structure 24. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 22 and Lane 4: Structure 24. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 4 c, which gave rise to − 5.48 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 4 e, which gave rise to − 5.19 as its mean value of ΔLk .

    Techniques Used: Agarose Gel Electrophoresis, Molecular Weight

    3) Product Images from "A System for the Analysis of BKV Non-coding Control Regions: Application to Clinical Isolates from an HIV/AIDS Patient"

    Article Title: A System for the Analysis of BKV Non-coding Control Regions: Application to Clinical Isolates from an HIV/AIDS Patient

    Journal: Virology

    doi: 10.1016/j.virol.2010.08.032

    The NCCR determines replication efficiency of BKV. (A) Schematic of the swap genome with the archetype virus (Dik) NCCR. SpeI and SacII sites were inserted into the pBR322-Dunlop or –Dik vectors flanking the majority of the NCCR and a PmlI site was inserted between the early and late regions. (B) RPTE cells were transfected with recircularized viral genome and low molecular weight DNA was harvested 5 dpt. Samples were linearized, digested with DpnI, and analyzed by Southern blotting. The left panel shows Dik and Dunlop wt replication compared to the swap vectors containing the three inserted restriction enzyme sites, Dik3 and Dun3 respectively. The right panel shows all possible swap combinations. Each construct is designated by a three letter abbreviation. A=archetype and R=rearranged. The first letter denotes the NCCR; second letter, early region; third letter, late region. Marker, HindIII digest of pGEM-TU; Mock, mock transfection.
    Figure Legend Snippet: The NCCR determines replication efficiency of BKV. (A) Schematic of the swap genome with the archetype virus (Dik) NCCR. SpeI and SacII sites were inserted into the pBR322-Dunlop or –Dik vectors flanking the majority of the NCCR and a PmlI site was inserted between the early and late regions. (B) RPTE cells were transfected with recircularized viral genome and low molecular weight DNA was harvested 5 dpt. Samples were linearized, digested with DpnI, and analyzed by Southern blotting. The left panel shows Dik and Dunlop wt replication compared to the swap vectors containing the three inserted restriction enzyme sites, Dik3 and Dun3 respectively. The right panel shows all possible swap combinations. Each construct is designated by a three letter abbreviation. A=archetype and R=rearranged. The first letter denotes the NCCR; second letter, early region; third letter, late region. Marker, HindIII digest of pGEM-TU; Mock, mock transfection.

    Techniques Used: Transfection, Molecular Weight, Southern Blot, Construct, Marker

    Related Articles

    Clone Assay:

    Article Title: Molecular basis for the structural diversity in serogroup O2-antigen polysaccharides in Klebsiella pneumoniae
    Article Snippet: .. Recombinant plasmids used in this study were constructed by cloning PCR fragments into the vector pBR322 by Gibson Assembly (New England Biolabs). .. Briefly, pBR322 was digested with the restriction endonucleases BamHI and SalI (New England Biolabs), and inserts were incorporated downstream of the tetracycline promoter by homologous recombination, mediated by primer sequences homologous to DNA flanking the restriction sites in pBR322.

    Amplification:

    Article Title: qnrD, a Novel Gene Conferring Transferable Quinolone Resistance in Salmonella enterica Serovar Kentucky and Bovismorbificans Strains of Human Origin ▿
    Article Snippet: .. The amplified fragment digested with EcoRV and SalI (New England BioLabs, Hitchin, United Kingdom) was then ligated into the tetA gene of vector pBR322 (New England Biolabs, Hitchin, United Kingdom) downstream of the tetA promoter digested with EcoRV and SalI (New England BioLabs, Hitchin, United Kingdom). .. The ligation product was electroporated into competent E. coli DH10B cells (Invitrogen, Cergy Pontoise, France).

    Electrophoretic Mobility Shift Assay:

    Article Title: Investigation into the antimicrobial action and mechanism of a novel endogenous peptide β-casein 197 from human milk
    Article Snippet: .. DNA binding assay Gel-retardation experiments were performed using 5 µL of a 25 µg mL−1 pBR322 vector from E. coli (BioLabs, New England). .. Plasmid DNA (pDNA) was exposed to 5 µL of different concentrations of β-casein 197 at 37 °C for 1 h prior to gel electrophoresis of the reaction mixtures through a 0.7% agarose gel in Tris–acetate EDTA buffer.

    DNA Binding Assay:

    Article Title: Investigation into the antimicrobial action and mechanism of a novel endogenous peptide β-casein 197 from human milk
    Article Snippet: .. DNA binding assay Gel-retardation experiments were performed using 5 µL of a 25 µg mL−1 pBR322 vector from E. coli (BioLabs, New England). .. Plasmid DNA (pDNA) was exposed to 5 µL of different concentrations of β-casein 197 at 37 °C for 1 h prior to gel electrophoresis of the reaction mixtures through a 0.7% agarose gel in Tris–acetate EDTA buffer.

    Concentration Assay:

    Article Title: A System for the Analysis of BKV Non-coding Control Regions: Application to Clinical Isolates from an HIV/AIDS Patient
    Article Snippet: .. The viral genome was excised from the pBR322 vector by BamHI (New England Biolabs, Ipswich, MA) digestion and then recirularized at a concentration of 10 ng/μL using T4 DNA ligase (New England Biolabs, Ipswich, MA). .. RPTE cells were seeded into 12 well plates and transfected with 0.6 μg DNA at 60–70% confluency using TransIT LT-1 transfection reagent (Mirus Bio, Madison, WI) according to the manufacturer’s instructions with a DNA to transfection reagent ratio of 1:6.

    Construct:

    Article Title: Molecular basis for the structural diversity in serogroup O2-antigen polysaccharides in Klebsiella pneumoniae
    Article Snippet: .. Recombinant plasmids used in this study were constructed by cloning PCR fragments into the vector pBR322 by Gibson Assembly (New England Biolabs). .. Briefly, pBR322 was digested with the restriction endonucleases BamHI and SalI (New England Biolabs), and inserts were incorporated downstream of the tetracycline promoter by homologous recombination, mediated by primer sequences homologous to DNA flanking the restriction sites in pBR322.

    Polymerase Chain Reaction:

    Article Title: Molecular basis for the structural diversity in serogroup O2-antigen polysaccharides in Klebsiella pneumoniae
    Article Snippet: .. Recombinant plasmids used in this study were constructed by cloning PCR fragments into the vector pBR322 by Gibson Assembly (New England Biolabs). .. Briefly, pBR322 was digested with the restriction endonucleases BamHI and SalI (New England Biolabs), and inserts were incorporated downstream of the tetracycline promoter by homologous recombination, mediated by primer sequences homologous to DNA flanking the restriction sites in pBR322.

    Article Title: Genetic Fusions of Heat-Labile Toxoid (LT) and Heat-Stable Toxin b (STb) of Porcine Enterotoxigenic Escherichia coli Elicit Protective Anti-LT and Anti-STb Antibodies ▿
    Article Snippet: .. Final PCR products (inserts) and vector pBR322 were digested with NheI and EagI restriction enzymes (New England Biolabs, Ipswich, MA) and ligated with T4 DNA ligase (Invitrogen) under standard conditions ( ). .. Ligated products were introduced into 1836-2 and TOP10 competent cells with standard electroporation ( ).

    Recombinant:

    Article Title: Molecular basis for the structural diversity in serogroup O2-antigen polysaccharides in Klebsiella pneumoniae
    Article Snippet: .. Recombinant plasmids used in this study were constructed by cloning PCR fragments into the vector pBR322 by Gibson Assembly (New England Biolabs). .. Briefly, pBR322 was digested with the restriction endonucleases BamHI and SalI (New England Biolabs), and inserts were incorporated downstream of the tetracycline promoter by homologous recombination, mediated by primer sequences homologous to DNA flanking the restriction sites in pBR322.

    Article Title: Dataset on the effects of spermidine on linking number differences between histone H1-free and histone H1-bound circular polynucleosomes
    Article Snippet: .. 2.4 Materials Recombinant human histone H2A/H2B dimer, histone H3.1/H4 tetramer, histone H10 , pBR322 vector, Nt.BspQI nicking endonuclease, T4 DNA ligase and Proteinase K were purchased from New England Biolabs Inc. (Singapore). .. QIAquick PCR Purification Kit was obtained from QIAGEN Singapore Pte.

    Plasmid Preparation:

    Article Title: NANOG Reporter Cell Lines Generated by Gene Targeting in Human Embryonic Stem Cells
    Article Snippet: .. The retrieval vector pBR322 was obtained from New England Biolabs. ..

    Article Title: Rational design of an orthogonal tryptophanyl nonsense suppressor tRNA
    Article Snippet: .. All restriction enzymes, T4 DNA ligase and vector pBR322 were from New England Biolabs (Ipswich, MA, USA). .. Vector pACYCDuet-1 was obtained from Novagen (San Diego, CA, USA).

    Article Title: A System for the Analysis of BKV Non-coding Control Regions: Application to Clinical Isolates from an HIV/AIDS Patient
    Article Snippet: .. The viral genome was excised from the pBR322 vector by BamHI (New England Biolabs, Ipswich, MA) digestion and then recirularized at a concentration of 10 ng/μL using T4 DNA ligase (New England Biolabs, Ipswich, MA). .. RPTE cells were seeded into 12 well plates and transfected with 0.6 μg DNA at 60–70% confluency using TransIT LT-1 transfection reagent (Mirus Bio, Madison, WI) according to the manufacturer’s instructions with a DNA to transfection reagent ratio of 1:6.

    Article Title: Molecular basis for the structural diversity in serogroup O2-antigen polysaccharides in Klebsiella pneumoniae
    Article Snippet: .. Recombinant plasmids used in this study were constructed by cloning PCR fragments into the vector pBR322 by Gibson Assembly (New England Biolabs). .. Briefly, pBR322 was digested with the restriction endonucleases BamHI and SalI (New England Biolabs), and inserts were incorporated downstream of the tetracycline promoter by homologous recombination, mediated by primer sequences homologous to DNA flanking the restriction sites in pBR322.

    Article Title: Genetic Fusions of Heat-Labile Toxoid (LT) and Heat-Stable Toxin b (STb) of Porcine Enterotoxigenic Escherichia coli Elicit Protective Anti-LT and Anti-STb Antibodies ▿
    Article Snippet: .. Final PCR products (inserts) and vector pBR322 were digested with NheI and EagI restriction enzymes (New England Biolabs, Ipswich, MA) and ligated with T4 DNA ligase (Invitrogen) under standard conditions ( ). .. Ligated products were introduced into 1836-2 and TOP10 competent cells with standard electroporation ( ).

    Article Title: Investigation into the antimicrobial action and mechanism of a novel endogenous peptide β-casein 197 from human milk
    Article Snippet: .. DNA binding assay Gel-retardation experiments were performed using 5 µL of a 25 µg mL−1 pBR322 vector from E. coli (BioLabs, New England). .. Plasmid DNA (pDNA) was exposed to 5 µL of different concentrations of β-casein 197 at 37 °C for 1 h prior to gel electrophoresis of the reaction mixtures through a 0.7% agarose gel in Tris–acetate EDTA buffer.

    Article Title: qnrD, a Novel Gene Conferring Transferable Quinolone Resistance in Salmonella enterica Serovar Kentucky and Bovismorbificans Strains of Human Origin ▿
    Article Snippet: .. The amplified fragment digested with EcoRV and SalI (New England BioLabs, Hitchin, United Kingdom) was then ligated into the tetA gene of vector pBR322 (New England Biolabs, Hitchin, United Kingdom) downstream of the tetA promoter digested with EcoRV and SalI (New England BioLabs, Hitchin, United Kingdom). .. The ligation product was electroporated into competent E. coli DH10B cells (Invitrogen, Cergy Pontoise, France).

    Article Title: Dataset on the effects of spermidine on linking number differences between histone H1-free and histone H1-bound circular polynucleosomes
    Article Snippet: .. 2.4 Materials Recombinant human histone H2A/H2B dimer, histone H3.1/H4 tetramer, histone H10 , pBR322 vector, Nt.BspQI nicking endonuclease, T4 DNA ligase and Proteinase K were purchased from New England Biolabs Inc. (Singapore). .. QIAquick PCR Purification Kit was obtained from QIAGEN Singapore Pte.

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    New England Biolabs pbr322 vector
    The DNA-binding properties of β-Casein 197. a ProtParam analysis of predicted binding residues of β-casein 197, predicted binding residues was labeled with a red “+”. b Gel retardation assays. Binding was assayed by the migration of pDNA. Various concentrations of peptides were incubated with 5 µL of 25 µg mL −1 <t>pBR322</t> vector from E. coli at 37 °C for 1 h, and then the reaction mixtures were applied to 0.7% agarose gel electrophoresis. Lane M DNA marker DL 10,000; Lane 1 5 µL pDNA as control; Lanes 2–6 mixture of pDNA and various concentrations of peptides (12.5, 25, 50, 100 and 500 µg mL −1 )
    Pbr322 Vector, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbr322 vector/product/New England Biolabs
    Average 95 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    pbr322 vector - by Bioz Stars, 2020-07
    95/100 stars
      Buy from Supplier

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    The DNA-binding properties of β-Casein 197. a ProtParam analysis of predicted binding residues of β-casein 197, predicted binding residues was labeled with a red “+”. b Gel retardation assays. Binding was assayed by the migration of pDNA. Various concentrations of peptides were incubated with 5 µL of 25 µg mL −1 pBR322 vector from E. coli at 37 °C for 1 h, and then the reaction mixtures were applied to 0.7% agarose gel electrophoresis. Lane M DNA marker DL 10,000; Lane 1 5 µL pDNA as control; Lanes 2–6 mixture of pDNA and various concentrations of peptides (12.5, 25, 50, 100 and 500 µg mL −1 )

    Journal: AMB Express

    Article Title: Investigation into the antimicrobial action and mechanism of a novel endogenous peptide β-casein 197 from human milk

    doi: 10.1186/s13568-017-0409-y

    Figure Lengend Snippet: The DNA-binding properties of β-Casein 197. a ProtParam analysis of predicted binding residues of β-casein 197, predicted binding residues was labeled with a red “+”. b Gel retardation assays. Binding was assayed by the migration of pDNA. Various concentrations of peptides were incubated with 5 µL of 25 µg mL −1 pBR322 vector from E. coli at 37 °C for 1 h, and then the reaction mixtures were applied to 0.7% agarose gel electrophoresis. Lane M DNA marker DL 10,000; Lane 1 5 µL pDNA as control; Lanes 2–6 mixture of pDNA and various concentrations of peptides (12.5, 25, 50, 100 and 500 µg mL −1 )

    Article Snippet: DNA binding assay Gel-retardation experiments were performed using 5 µL of a 25 µg mL−1 pBR322 vector from E. coli (BioLabs, New England).

    Techniques: Binding Assay, Labeling, Electrophoretic Mobility Shift Assay, Migration, Incubation, Plasmid Preparation, Agarose Gel Electrophoresis, Marker

    GM1-ELISA to measure binding of LT 192 and LT 192 -Gly:Pro-STb proteins to GM1. Total proteins extracted from equivalent amounts of cells (determined by OD readings) of each strain (3030-2 [K88ac + LT + STb + ], 8017 [1836-2/pBR322], 8035 [1836-2 LT], 8221 [1836-2 LT 192 ], 8145 [1836-2 LT-Gly:Pro-STb], and 8488 [1836-2 LT 192 -Gly:Pro-STb]) were tested in GM1 binding using anti-CT as the primary antibody (1:5,000) and horseradish peroxidase-conjugated goat anti-rabbit IgG (1:5,000) as the secondary antibody. Optical densities, measured at 405 nm, showed significant differences for 3030-2, 8035, and 8221 strains ( P

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Genetic Fusions of Heat-Labile Toxoid (LT) and Heat-Stable Toxin b (STb) of Porcine Enterotoxigenic Escherichia coli Elicit Protective Anti-LT and Anti-STb Antibodies ▿

    doi: 10.1128/CVI.00095-10

    Figure Lengend Snippet: GM1-ELISA to measure binding of LT 192 and LT 192 -Gly:Pro-STb proteins to GM1. Total proteins extracted from equivalent amounts of cells (determined by OD readings) of each strain (3030-2 [K88ac + LT + STb + ], 8017 [1836-2/pBR322], 8035 [1836-2 LT], 8221 [1836-2 LT 192 ], 8145 [1836-2 LT-Gly:Pro-STb], and 8488 [1836-2 LT 192 -Gly:Pro-STb]) were tested in GM1 binding using anti-CT as the primary antibody (1:5,000) and horseradish peroxidase-conjugated goat anti-rabbit IgG (1:5,000) as the secondary antibody. Optical densities, measured at 405 nm, showed significant differences for 3030-2, 8035, and 8221 strains ( P

    Article Snippet: Final PCR products (inserts) and vector pBR322 were digested with NheI and EagI restriction enzymes (New England Biolabs, Ipswich, MA) and ligated with T4 DNA ligase (Invitrogen) under standard conditions ( ).

    Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay

    Detection of toxic activity in a porcine ligated-gut-loop assay. Individual ligated intestinal loops from the ileum and jejunum sections were inoculated with 2 × 10 9 CFU of overnight culture from strain 3030-2 (K88 + LT + STb + , a positive control), 8488 (1836-2 LT 192 -Gly:Pro-STb), 8221(1836-2 LT 192 ), 8017 (1836-2/pBR322, a negative control), or 8816 (1836-2 STb). Fluid accumulation was measured after 8 h postinoculation; the data are presented in the inserted table. Statistical analysis indicated that the fluid accumulation in loops incubated with strains 8488 and 8221 was not significantly different from that in loops incubated with the negative control 8017 strain ( P > 0.05), whereas fluid accumulated in the loops incubated with 8816 and 3030-2 was significantly different ( P

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Genetic Fusions of Heat-Labile Toxoid (LT) and Heat-Stable Toxin b (STb) of Porcine Enterotoxigenic Escherichia coli Elicit Protective Anti-LT and Anti-STb Antibodies ▿

    doi: 10.1128/CVI.00095-10

    Figure Lengend Snippet: Detection of toxic activity in a porcine ligated-gut-loop assay. Individual ligated intestinal loops from the ileum and jejunum sections were inoculated with 2 × 10 9 CFU of overnight culture from strain 3030-2 (K88 + LT + STb + , a positive control), 8488 (1836-2 LT 192 -Gly:Pro-STb), 8221(1836-2 LT 192 ), 8017 (1836-2/pBR322, a negative control), or 8816 (1836-2 STb). Fluid accumulation was measured after 8 h postinoculation; the data are presented in the inserted table. Statistical analysis indicated that the fluid accumulation in loops incubated with strains 8488 and 8221 was not significantly different from that in loops incubated with the negative control 8017 strain ( P > 0.05), whereas fluid accumulated in the loops incubated with 8816 and 3030-2 was significantly different ( P

    Article Snippet: Final PCR products (inserts) and vector pBR322 were digested with NheI and EagI restriction enzymes (New England Biolabs, Ipswich, MA) and ligated with T4 DNA ligase (Invitrogen) under standard conditions ( ).

    Techniques: Activity Assay, Positive Control, Negative Control, Incubation

    (a) Illustration of our preparations of Structure 26 and Structure 28 in the presence of 5 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 26 and Structure 28. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 26 and Lane 4: Structure 28. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 5 c, which gave rise to − 5.20 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 5 e, which gave rise to − 5.32 as its mean value of ΔLk .

    Journal: Data in Brief

    Article Title: Dataset on the effects of spermidine on linking number differences between histone H1-free and histone H1-bound circular polynucleosomes

    doi: 10.1016/j.dib.2018.01.091

    Figure Lengend Snippet: (a) Illustration of our preparations of Structure 26 and Structure 28 in the presence of 5 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 26 and Structure 28. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 26 and Lane 4: Structure 28. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 5 c, which gave rise to − 5.20 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 5 e, which gave rise to − 5.32 as its mean value of ΔLk .

    Article Snippet: 2.4 Materials Recombinant human histone H2A/H2B dimer, histone H3.1/H4 tetramer, histone H10 , pBR322 vector, Nt.BspQI nicking endonuclease, T4 DNA ligase and Proteinase K were purchased from New England Biolabs Inc. (Singapore).

    Techniques: Agarose Gel Electrophoresis, Molecular Weight

    (a) Illustration of our preparations of Structure 10 and Structure 12 in the presence of 0.75 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 10 and Structure 12. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 10 and Lane 4: Structure 12. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 1 c, which gave rise to − 6.34 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 1 e, which gave rise to − 6.07 as its mean value of ΔLk .

    Journal: Data in Brief

    Article Title: Dataset on the effects of spermidine on linking number differences between histone H1-free and histone H1-bound circular polynucleosomes

    doi: 10.1016/j.dib.2018.01.091

    Figure Lengend Snippet: (a) Illustration of our preparations of Structure 10 and Structure 12 in the presence of 0.75 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 10 and Structure 12. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 10 and Lane 4: Structure 12. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 1 c, which gave rise to − 6.34 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 1 e, which gave rise to − 6.07 as its mean value of ΔLk .

    Article Snippet: 2.4 Materials Recombinant human histone H2A/H2B dimer, histone H3.1/H4 tetramer, histone H10 , pBR322 vector, Nt.BspQI nicking endonuclease, T4 DNA ligase and Proteinase K were purchased from New England Biolabs Inc. (Singapore).

    Techniques: Agarose Gel Electrophoresis, Molecular Weight

    (a) Illustration of our preparations of Structure 18 and Structure 20 in the presence of 2 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 18 and Structure 20. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 18 and Lane 4: Structure 20. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 3 c, which gave rise to − 6.41 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 3 e, which gave rise to − 5.98 as its mean value of ΔLk .

    Journal: Data in Brief

    Article Title: Dataset on the effects of spermidine on linking number differences between histone H1-free and histone H1-bound circular polynucleosomes

    doi: 10.1016/j.dib.2018.01.091

    Figure Lengend Snippet: (a) Illustration of our preparations of Structure 18 and Structure 20 in the presence of 2 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 18 and Structure 20. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 18 and Lane 4: Structure 20. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 3 c, which gave rise to − 6.41 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 3 e, which gave rise to − 5.98 as its mean value of ΔLk .

    Article Snippet: 2.4 Materials Recombinant human histone H2A/H2B dimer, histone H3.1/H4 tetramer, histone H10 , pBR322 vector, Nt.BspQI nicking endonuclease, T4 DNA ligase and Proteinase K were purchased from New England Biolabs Inc. (Singapore).

    Techniques: Agarose Gel Electrophoresis, Molecular Weight

    (a) Illustration of our preparations of Structure 14 and Structure 16 in the presence of 0 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 14 and Structure 16. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 14 and Lane 4: Structure 16. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 2 c, which gave rise to − 6.01 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 2 e, which gave rise to − 5.98 as its mean value of ΔLk .

    Journal: Data in Brief

    Article Title: Dataset on the effects of spermidine on linking number differences between histone H1-free and histone H1-bound circular polynucleosomes

    doi: 10.1016/j.dib.2018.01.091

    Figure Lengend Snippet: (a) Illustration of our preparations of Structure 14 and Structure 16 in the presence of 0 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 14 and Structure 16. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 14 and Lane 4: Structure 16. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 2 c, which gave rise to − 6.01 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 2 e, which gave rise to − 5.98 as its mean value of ΔLk .

    Article Snippet: 2.4 Materials Recombinant human histone H2A/H2B dimer, histone H3.1/H4 tetramer, histone H10 , pBR322 vector, Nt.BspQI nicking endonuclease, T4 DNA ligase and Proteinase K were purchased from New England Biolabs Inc. (Singapore).

    Techniques: Agarose Gel Electrophoresis, Molecular Weight

    (a) Illustration of our preparations of Structure 22 and Structure 24 in the presence of 3 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 22 and Structure 24. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 22 and Lane 4: Structure 24. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 4 c, which gave rise to − 5.48 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 4 e, which gave rise to − 5.19 as its mean value of ΔLk .

    Journal: Data in Brief

    Article Title: Dataset on the effects of spermidine on linking number differences between histone H1-free and histone H1-bound circular polynucleosomes

    doi: 10.1016/j.dib.2018.01.091

    Figure Lengend Snippet: (a) Illustration of our preparations of Structure 22 and Structure 24 in the presence of 3 mM spermidine. (b) Chloroquine-based agarose gel electrophoretic analysis on Structure 22 and Structure 24. Lane 1: molecular weight markers; Lane 2: relaxed forms of pBR322; Lane 3: Structure 22 and Lane 4: Structure 24. (c) Densitometry tracing of gel electrophoretic results in Lane 3. (d) Plot of Gauss fit on the data shown in Fig. 4 c, which gave rise to − 5.48 as its mean value of ΔLk (e) Densitometry tracing of gel electrophoretic results in Lane 4. (f) Plot of Gauss fit on the data shown in Fig. 4 e, which gave rise to − 5.19 as its mean value of ΔLk .

    Article Snippet: 2.4 Materials Recombinant human histone H2A/H2B dimer, histone H3.1/H4 tetramer, histone H10 , pBR322 vector, Nt.BspQI nicking endonuclease, T4 DNA ligase and Proteinase K were purchased from New England Biolabs Inc. (Singapore).

    Techniques: Agarose Gel Electrophoresis, Molecular Weight

    The NCCR determines replication efficiency of BKV. (A) Schematic of the swap genome with the archetype virus (Dik) NCCR. SpeI and SacII sites were inserted into the pBR322-Dunlop or –Dik vectors flanking the majority of the NCCR and a PmlI site was inserted between the early and late regions. (B) RPTE cells were transfected with recircularized viral genome and low molecular weight DNA was harvested 5 dpt. Samples were linearized, digested with DpnI, and analyzed by Southern blotting. The left panel shows Dik and Dunlop wt replication compared to the swap vectors containing the three inserted restriction enzyme sites, Dik3 and Dun3 respectively. The right panel shows all possible swap combinations. Each construct is designated by a three letter abbreviation. A=archetype and R=rearranged. The first letter denotes the NCCR; second letter, early region; third letter, late region. Marker, HindIII digest of pGEM-TU; Mock, mock transfection.

    Journal: Virology

    Article Title: A System for the Analysis of BKV Non-coding Control Regions: Application to Clinical Isolates from an HIV/AIDS Patient

    doi: 10.1016/j.virol.2010.08.032

    Figure Lengend Snippet: The NCCR determines replication efficiency of BKV. (A) Schematic of the swap genome with the archetype virus (Dik) NCCR. SpeI and SacII sites were inserted into the pBR322-Dunlop or –Dik vectors flanking the majority of the NCCR and a PmlI site was inserted between the early and late regions. (B) RPTE cells were transfected with recircularized viral genome and low molecular weight DNA was harvested 5 dpt. Samples were linearized, digested with DpnI, and analyzed by Southern blotting. The left panel shows Dik and Dunlop wt replication compared to the swap vectors containing the three inserted restriction enzyme sites, Dik3 and Dun3 respectively. The right panel shows all possible swap combinations. Each construct is designated by a three letter abbreviation. A=archetype and R=rearranged. The first letter denotes the NCCR; second letter, early region; third letter, late region. Marker, HindIII digest of pGEM-TU; Mock, mock transfection.

    Article Snippet: The viral genome was excised from the pBR322 vector by BamHI (New England Biolabs, Ipswich, MA) digestion and then recirularized at a concentration of 10 ng/μL using T4 DNA ligase (New England Biolabs, Ipswich, MA).

    Techniques: Transfection, Molecular Weight, Southern Blot, Construct, Marker