pet 28a vector  (Millipore)


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    Structured Review

    Millipore pet 28a vector
    Pet 28a Vector, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 513 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pet 28a vector/product/Millipore
    Average 99 stars, based on 513 article reviews
    Price from $9.99 to $1999.99
    pet 28a vector - by Bioz Stars, 2020-02
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Molecular Characterization and Identification of Target Protein of an Important Vesicle Trafficking Gene AlRab7 from a Salt Excreting Halophyte Aeluropus lagopoides
    Article Snippet: .. The amplified product was gel purified, digested, and cloned in pET 28a expression vector (Novagen). .. The recombinant plasmid was transformed in BL21 (DE3) star Escherichia coli strain and grown in Luria Bertani (LB) medium at 37°C to OD600 0.5.

    Article Title: Tailoring the properties of (catalytically)-active inclusion bodies
    Article Snippet: .. If not stated otherwise, all gene fusions consisted of gene fragments coding for a coiled-coil domain (here TDoT or 3HAMP), a linker polypeptide, consisting of a protease Factor Xa cleavage site and a triple (GGGS)3 and the respective target proteins/enzymes cloned into a pET-28a vector (Novagen, Merck KGaA, Frankfurt, Germany). ..

    Article Title: Transglutaminase 2-specific autoantibodies in celiac disease target clustered, N-terminal epitopes not displayed on the surface of cells 1
    Article Snippet: .. Mouse TG2 was obtained by extraction of RNA from mouse liver biopsies and cloning of TG2-encoding cDNA into the pET-28a vector (Novagen) as described for human TG2 ( ). .. Mouse TG2 and human TG2 mutants were produced in E. coli in the same way as wild-type human TG2.

    Article Title: 1H, 13C, and 15N resonance assignments and secondary structure prediction of the full-length transition state regulator AbrB from Bacillus anthracis
    Article Snippet: .. Full length AbrB (residues 1–94) from B. anthracis was cloned into expression vector pET-28a (Novagen) with a Thrombin cleavable N-terminal histidine tag and transformed into BL21(DE3) cells (Genesee) for expression. ..

    Article Title: Hfq, a Novel Pleiotropic Regulator of Virulence-Associated Genes in Francisella tularensis ▿
    Article Snippet: .. The hfq gene of LVS was cloned into the pET-28a expression vector (Novagen) to create a protein with a histidine tag at the N terminus. ..

    Article Title: QnrS1 structure–activity relationships
    Article Snippet: .. The resulting DNA fragment was double-digested with SacI and XhoI (New England Biolabs, Ipswich, MA, USA) and cloned into expression vector pET-28a (Novagen, USA). .. Plasmid pET-28a:QnrS1 was transformed into E. coli BL21(DE3) (New England Biolabs) and plated on Luria–Bertani (LB) agar (Becton Dickinson, USA) containing 100 mg/L kanamycin to maintain the plasmid.

    Article Title: Bmserpin2 Is Involved in BmNPV Infection by Suppressing Melanization in Bombyx mori
    Article Snippet: .. Recombinant Bmserpin2 and Antibody Preparation The cloned cDNA fragment that contained mature Bmserpin2 ligated into the expression vector pET-28a (Novagen) with restriction enzyme sites BamH1 and Xho1 , and the insertion was confirmed by DNA sequencing in the recombinant BmSerpin2-pET-28a plasmid and then the recombinant plasmid was transformed into Escherichia coli BL21 (Novagen) competent cells which were grown in fresh Luria-Bertani (LB) medium. .. The positive plasmid was induced by isopropyl β-D-thiogalactoside (IPTG) (1 mM) to induce recombinant Bmserpin2 expression for 6 h at 37 °C.

    Centrifugation:

    Article Title: 1H, 13C, and 15N resonance assignments and secondary structure prediction of the full-length transition state regulator AbrB from Bacillus anthracis
    Article Snippet: Full length AbrB (residues 1–94) from B. anthracis was cloned into expression vector pET-28a (Novagen) with a Thrombin cleavable N-terminal histidine tag and transformed into BL21(DE3) cells (Genesee) for expression. .. The cells were harvested by centrifugation 4 h postinduction at 7,0009×g for 15 min.

    Article Title: Differential regulation of dopaminergic gene expression by Er81
    Article Snippet: ER81 used in the EMSA experiments was generated by over-expression in recombinant pLysS E. coli bacteria containing a pET-28A expression plasmid (Novagen) containing an Er81 cDNA isolated from mouse OB RNA using the Oligotex mRNA mini-kit (Qiagen). .. Cells were pelleted by centrifugation and then resuspended in 30 ml Tris buffered saline (pH 7.2).

    Article Title: Bmserpin2 Is Involved in BmNPV Infection by Suppressing Melanization in Bombyx mori
    Article Snippet: Recombinant Bmserpin2 and Antibody Preparation The cloned cDNA fragment that contained mature Bmserpin2 ligated into the expression vector pET-28a (Novagen) with restriction enzyme sites BamH1 and Xho1 , and the insertion was confirmed by DNA sequencing in the recombinant BmSerpin2-pET-28a plasmid and then the recombinant plasmid was transformed into Escherichia coli BL21 (Novagen) competent cells which were grown in fresh Luria-Bertani (LB) medium. .. After centrifugation at 7500× g for 10 min at 4 °C, bacteria were collected and sonicated in phosphate-buffered saline (PBS).

    Amplification:

    Article Title: Molecular Characterization and Identification of Target Protein of an Important Vesicle Trafficking Gene AlRab7 from a Salt Excreting Halophyte Aeluropus lagopoides
    Article Snippet: .. The amplified product was gel purified, digested, and cloned in pET 28a expression vector (Novagen). .. The recombinant plasmid was transformed in BL21 (DE3) star Escherichia coli strain and grown in Luria Bertani (LB) medium at 37°C to OD600 0.5.

    Article Title: QnrS1 structure–activity relationships
    Article Snippet: A DNA fragment encompassing the qnrS1 gene was amplified by PCR from plasmid pMG306 (GenBank accession no. ) using the primers listed in Table . .. The resulting DNA fragment was double-digested with SacI and XhoI (New England Biolabs, Ipswich, MA, USA) and cloned into expression vector pET-28a (Novagen, USA).

    Article Title: Crystallization and preliminary X-ray crystallographic studies of fatty acid-CoA racemase from Mycobacterium tuberculosis H37Rv
    Article Snippet: The far gene of M. tuberculosis H37Rv was amplified by polymerase chain reaction from the bacterial genomic DNA using Pyrobest (Takara) and two primers (5′-GGAATTC CATATG ATGACGACCGGGGGGC-3′ and 5′-CCG CTCGAG TCATTACCAGTTGATTTCGTCGATC-3′). .. The PCR product was purified, digested with Nde I and Xho I and ligated into the pET-28a vector (Novagen).

    Construct:

    Article Title: Molecular Characterization, Tissue Distribution, Subcellular Localization and Actin-Sequestering Function of a Thymosin Protein from Silkworm
    Article Snippet: .. Construction of Recombinant Plasmids We used pHelix-BmTHY constructed by our laboratory as a template to amplify BmTHY ORF by PCR, with primers complementary to the flanking sequences of the ORF with Eco RI and Xho I recognition sites as follows: P1 : 5′- CGC GAATT CATGGCCTGCTCC -3′ ; P2 : 5′- CCG CTCGAG TCAAGCTGATTTCTCTT -3′ ; The PCR products were purified after electrophoresis on 1% agarose gel using the PCR Rapid Purification Kit (BioDev-Tech, China).After digestion with Eco RI and Xho I, the purified PCR products were subcloned into the expression vector pET-28a (Novagen, Darmstadt, Germany) using T4 DNA ligase (Promega, USA) and transformed into E. coli TG1 cells (maintained in our laboratory) for screening purposes. .. A positive colony with BmTHY gene in the plasmid was identified by double digestion of the plasmid, followed by analysis on 1% agarose gel electrophoresis and was subsequently verified by DNA sequencing.

    Article Title: Detection of anti-preS1 antibodies for recovery of hepatitis B patients by immunoassay
    Article Snippet: Plasmids, E. coli strain The plasmid pADR-1 containing HBV genome (adr subtype) was constructed by Wu et al[ ]. .. Expression vector pET-28a (+) was purchased from Novagen.

    Electrophoresis:

    Article Title: Molecular Characterization, Tissue Distribution, Subcellular Localization and Actin-Sequestering Function of a Thymosin Protein from Silkworm
    Article Snippet: .. Construction of Recombinant Plasmids We used pHelix-BmTHY constructed by our laboratory as a template to amplify BmTHY ORF by PCR, with primers complementary to the flanking sequences of the ORF with Eco RI and Xho I recognition sites as follows: P1 : 5′- CGC GAATT CATGGCCTGCTCC -3′ ; P2 : 5′- CCG CTCGAG TCAAGCTGATTTCTCTT -3′ ; The PCR products were purified after electrophoresis on 1% agarose gel using the PCR Rapid Purification Kit (BioDev-Tech, China).After digestion with Eco RI and Xho I, the purified PCR products were subcloned into the expression vector pET-28a (Novagen, Darmstadt, Germany) using T4 DNA ligase (Promega, USA) and transformed into E. coli TG1 cells (maintained in our laboratory) for screening purposes. .. A positive colony with BmTHY gene in the plasmid was identified by double digestion of the plasmid, followed by analysis on 1% agarose gel electrophoresis and was subsequently verified by DNA sequencing.

    Incubation:

    Article Title: Hfq, a Novel Pleiotropic Regulator of Virulence-Associated Genes in Francisella tularensis ▿
    Article Snippet: The hfq gene of LVS was cloned into the pET-28a expression vector (Novagen) to create a protein with a histidine tag at the N terminus. .. Antiserum was purified by incubation with an F. tularensis Δ hfq bacterial lysate before it was used for immunoblotting.

    Expressing:

    Article Title: Molecular Characterization and Identification of Target Protein of an Important Vesicle Trafficking Gene AlRab7 from a Salt Excreting Halophyte Aeluropus lagopoides
    Article Snippet: .. The amplified product was gel purified, digested, and cloned in pET 28a expression vector (Novagen). .. The recombinant plasmid was transformed in BL21 (DE3) star Escherichia coli strain and grown in Luria Bertani (LB) medium at 37°C to OD600 0.5.

    Article Title: Tailoring the properties of (catalytically)-active inclusion bodies
    Article Snippet: Paragraph title: Construction of expression plasmids ... If not stated otherwise, all gene fusions consisted of gene fragments coding for a coiled-coil domain (here TDoT or 3HAMP), a linker polypeptide, consisting of a protease Factor Xa cleavage site and a triple (GGGS)3 and the respective target proteins/enzymes cloned into a pET-28a vector (Novagen, Merck KGaA, Frankfurt, Germany).

    Article Title: 1H, 13C, and 15N resonance assignments and secondary structure prediction of the full-length transition state regulator AbrB from Bacillus anthracis
    Article Snippet: .. Full length AbrB (residues 1–94) from B. anthracis was cloned into expression vector pET-28a (Novagen) with a Thrombin cleavable N-terminal histidine tag and transformed into BL21(DE3) cells (Genesee) for expression. ..

    Article Title: A broad pH range and processive chitinase from a metagenome library
    Article Snippet: .. E.coli BL21(DE3) and vector pET-28a(+) (Novagen, USA) were used as the recombinant protein expression system. .. Chemicals and enzymes FideliTaq PCR Master Mix (USB, USA) was used for DNA amplification.

    Article Title: Mechanism of protective immunity by vaccination with recombinant Echinococcus granulosus glutathione S-transferase (Chinese strain) in mice
    Article Snippet: .. The target fragment was purified using a gel cleanup kit (SBS Genetech, Co., Ltd.) and inserted between the Eco RI and Not I sites of the expression vector pET-28a (Novagen; Merck KGaA, Darmstadt, Germany). .. The recombinant vector EgGST/pET-28a was identified by restriction digestion using Eco RI and Not I and the EgGST insert was verified by sequencing, performed by Sangon Biotech Co., Ltd. (Shanghai, China).

    Article Title: Hfq, a Novel Pleiotropic Regulator of Virulence-Associated Genes in Francisella tularensis ▿
    Article Snippet: .. The hfq gene of LVS was cloned into the pET-28a expression vector (Novagen) to create a protein with a histidine tag at the N terminus. ..

    Article Title: High Level Expression and Purification of the Clinically Active Antimicrobial Peptide P-113 in Escherichia coli
    Article Snippet: .. The expression vector pET-28a and host BL21 (DE3) were purchased from Novagen (Merck Millipore, Burlington, MA, USA). .. Restriction enzymes and T4 ligase were purchased from Thermo Scientific (Waltham, MA, USA).

    Article Title: QnrS1 structure–activity relationships
    Article Snippet: .. The resulting DNA fragment was double-digested with SacI and XhoI (New England Biolabs, Ipswich, MA, USA) and cloned into expression vector pET-28a (Novagen, USA). .. Plasmid pET-28a:QnrS1 was transformed into E. coli BL21(DE3) (New England Biolabs) and plated on Luria–Bertani (LB) agar (Becton Dickinson, USA) containing 100 mg/L kanamycin to maintain the plasmid.

    Article Title: Crystallization and preliminary X-ray crystallographic studies of fatty acid-CoA racemase from Mycobacterium tuberculosis H37Rv
    Article Snippet: Paragraph title: 2.1. Expression and purification ... The PCR product was purified, digested with Nde I and Xho I and ligated into the pET-28a vector (Novagen).

    Article Title: Differential regulation of dopaminergic gene expression by Er81
    Article Snippet: .. ER81 used in the EMSA experiments was generated by over-expression in recombinant pLysS E. coli bacteria containing a pET-28A expression plasmid (Novagen) containing an Er81 cDNA isolated from mouse OB RNA using the Oligotex mRNA mini-kit (Qiagen). .. ER81 expression was induced with 1mMIPTG in bacterial cultures and harvested 6 hours after induction.

    Article Title: High Level Expression and Purification of the Clinically Active Antimicrobial Peptide P-113 in Escherichia coli
    Article Snippet: .. Materials and Methods The expression vector pET-28a and host BL21 (DE3) were purchased from Novagen (Merck Millipore, Burlington, MA, USA). .. Restriction enzymes and T4 ligase were purchased from Thermo Scientific (Waltham, MA, USA).

    Article Title: Bmserpin2 Is Involved in BmNPV Infection by Suppressing Melanization in Bombyx mori
    Article Snippet: .. Recombinant Bmserpin2 and Antibody Preparation The cloned cDNA fragment that contained mature Bmserpin2 ligated into the expression vector pET-28a (Novagen) with restriction enzyme sites BamH1 and Xho1 , and the insertion was confirmed by DNA sequencing in the recombinant BmSerpin2-pET-28a plasmid and then the recombinant plasmid was transformed into Escherichia coli BL21 (Novagen) competent cells which were grown in fresh Luria-Bertani (LB) medium. .. The positive plasmid was induced by isopropyl β-D-thiogalactoside (IPTG) (1 mM) to induce recombinant Bmserpin2 expression for 6 h at 37 °C.

    Article Title: Molecular Characterization, Tissue Distribution, Subcellular Localization and Actin-Sequestering Function of a Thymosin Protein from Silkworm
    Article Snippet: .. Construction of Recombinant Plasmids We used pHelix-BmTHY constructed by our laboratory as a template to amplify BmTHY ORF by PCR, with primers complementary to the flanking sequences of the ORF with Eco RI and Xho I recognition sites as follows: P1 : 5′- CGC GAATT CATGGCCTGCTCC -3′ ; P2 : 5′- CCG CTCGAG TCAAGCTGATTTCTCTT -3′ ; The PCR products were purified after electrophoresis on 1% agarose gel using the PCR Rapid Purification Kit (BioDev-Tech, China).After digestion with Eco RI and Xho I, the purified PCR products were subcloned into the expression vector pET-28a (Novagen, Darmstadt, Germany) using T4 DNA ligase (Promega, USA) and transformed into E. coli TG1 cells (maintained in our laboratory) for screening purposes. .. A positive colony with BmTHY gene in the plasmid was identified by double digestion of the plasmid, followed by analysis on 1% agarose gel electrophoresis and was subsequently verified by DNA sequencing.

    Article Title: Detection of anti-preS1 antibodies for recovery of hepatitis B patients by immunoassay
    Article Snippet: .. Expression vector pET-28a (+) was purchased from Novagen. ..

    Transformation Assay:

    Article Title: Molecular Characterization and Identification of Target Protein of an Important Vesicle Trafficking Gene AlRab7 from a Salt Excreting Halophyte Aeluropus lagopoides
    Article Snippet: The amplified product was gel purified, digested, and cloned in pET 28a expression vector (Novagen). .. The recombinant plasmid was transformed in BL21 (DE3) star Escherichia coli strain and grown in Luria Bertani (LB) medium at 37°C to OD600 0.5.

    Article Title: 1H, 13C, and 15N resonance assignments and secondary structure prediction of the full-length transition state regulator AbrB from Bacillus anthracis
    Article Snippet: .. Full length AbrB (residues 1–94) from B. anthracis was cloned into expression vector pET-28a (Novagen) with a Thrombin cleavable N-terminal histidine tag and transformed into BL21(DE3) cells (Genesee) for expression. ..

    Article Title: Mechanism of protective immunity by vaccination with recombinant Echinococcus granulosus glutathione S-transferase (Chinese strain) in mice
    Article Snippet: The target fragment was purified using a gel cleanup kit (SBS Genetech, Co., Ltd.) and inserted between the Eco RI and Not I sites of the expression vector pET-28a (Novagen; Merck KGaA, Darmstadt, Germany). .. E. coli BL21 (DE3) pLysS, provided by Dr Xiao Wei (University of Saskatchewan, Saskatoon, Canada) was transformed for induced expression of His6-tagged EgGST protein.

    Article Title: Bmserpin2 Is Involved in BmNPV Infection by Suppressing Melanization in Bombyx mori
    Article Snippet: .. Recombinant Bmserpin2 and Antibody Preparation The cloned cDNA fragment that contained mature Bmserpin2 ligated into the expression vector pET-28a (Novagen) with restriction enzyme sites BamH1 and Xho1 , and the insertion was confirmed by DNA sequencing in the recombinant BmSerpin2-pET-28a plasmid and then the recombinant plasmid was transformed into Escherichia coli BL21 (Novagen) competent cells which were grown in fresh Luria-Bertani (LB) medium. .. The positive plasmid was induced by isopropyl β-D-thiogalactoside (IPTG) (1 mM) to induce recombinant Bmserpin2 expression for 6 h at 37 °C.

    Article Title: Molecular Characterization, Tissue Distribution, Subcellular Localization and Actin-Sequestering Function of a Thymosin Protein from Silkworm
    Article Snippet: .. Construction of Recombinant Plasmids We used pHelix-BmTHY constructed by our laboratory as a template to amplify BmTHY ORF by PCR, with primers complementary to the flanking sequences of the ORF with Eco RI and Xho I recognition sites as follows: P1 : 5′- CGC GAATT CATGGCCTGCTCC -3′ ; P2 : 5′- CCG CTCGAG TCAAGCTGATTTCTCTT -3′ ; The PCR products were purified after electrophoresis on 1% agarose gel using the PCR Rapid Purification Kit (BioDev-Tech, China).After digestion with Eco RI and Xho I, the purified PCR products were subcloned into the expression vector pET-28a (Novagen, Darmstadt, Germany) using T4 DNA ligase (Promega, USA) and transformed into E. coli TG1 cells (maintained in our laboratory) for screening purposes. .. A positive colony with BmTHY gene in the plasmid was identified by double digestion of the plasmid, followed by analysis on 1% agarose gel electrophoresis and was subsequently verified by DNA sequencing.

    Over Expression:

    Article Title: Differential regulation of dopaminergic gene expression by Er81
    Article Snippet: .. ER81 used in the EMSA experiments was generated by over-expression in recombinant pLysS E. coli bacteria containing a pET-28A expression plasmid (Novagen) containing an Er81 cDNA isolated from mouse OB RNA using the Oligotex mRNA mini-kit (Qiagen). .. ER81 expression was induced with 1mMIPTG in bacterial cultures and harvested 6 hours after induction.

    Generated:

    Article Title: Differential regulation of dopaminergic gene expression by Er81
    Article Snippet: .. ER81 used in the EMSA experiments was generated by over-expression in recombinant pLysS E. coli bacteria containing a pET-28A expression plasmid (Novagen) containing an Er81 cDNA isolated from mouse OB RNA using the Oligotex mRNA mini-kit (Qiagen). .. ER81 expression was induced with 1mMIPTG in bacterial cultures and harvested 6 hours after induction.

    DNA Sequencing:

    Article Title: Bmserpin2 Is Involved in BmNPV Infection by Suppressing Melanization in Bombyx mori
    Article Snippet: .. Recombinant Bmserpin2 and Antibody Preparation The cloned cDNA fragment that contained mature Bmserpin2 ligated into the expression vector pET-28a (Novagen) with restriction enzyme sites BamH1 and Xho1 , and the insertion was confirmed by DNA sequencing in the recombinant BmSerpin2-pET-28a plasmid and then the recombinant plasmid was transformed into Escherichia coli BL21 (Novagen) competent cells which were grown in fresh Luria-Bertani (LB) medium. .. The positive plasmid was induced by isopropyl β-D-thiogalactoside (IPTG) (1 mM) to induce recombinant Bmserpin2 expression for 6 h at 37 °C.

    Article Title: Molecular Characterization, Tissue Distribution, Subcellular Localization and Actin-Sequestering Function of a Thymosin Protein from Silkworm
    Article Snippet: Construction of Recombinant Plasmids We used pHelix-BmTHY constructed by our laboratory as a template to amplify BmTHY ORF by PCR, with primers complementary to the flanking sequences of the ORF with Eco RI and Xho I recognition sites as follows: P1 : 5′- CGC GAATT CATGGCCTGCTCC -3′ ; P2 : 5′- CCG CTCGAG TCAAGCTGATTTCTCTT -3′ ; The PCR products were purified after electrophoresis on 1% agarose gel using the PCR Rapid Purification Kit (BioDev-Tech, China).After digestion with Eco RI and Xho I, the purified PCR products were subcloned into the expression vector pET-28a (Novagen, Darmstadt, Germany) using T4 DNA ligase (Promega, USA) and transformed into E. coli TG1 cells (maintained in our laboratory) for screening purposes. .. A positive colony with BmTHY gene in the plasmid was identified by double digestion of the plasmid, followed by analysis on 1% agarose gel electrophoresis and was subsequently verified by DNA sequencing.

    Sequencing:

    Article Title: Molecular Characterization and Identification of Target Protein of an Important Vesicle Trafficking Gene AlRab7 from a Salt Excreting Halophyte Aeluropus lagopoides
    Article Snippet: Using these primers the complete coding sequence for the AlRab7 was PCR amplified (150 ng of each primer, 200 μM dNTPs, 2.5 U Taq DNA polymerase in a 50 μL reaction, with 94°C, 5 min for 1 cycle, 94°C, 1 min, 55°C, 1 min, and 72°C, 1 min for 30 cycles; and 72°C, 7 min for 1 cycle) using Aeluropus cDNA clone as template. .. The amplified product was gel purified, digested, and cloned in pET 28a expression vector (Novagen).

    Article Title: Transglutaminase 2-specific autoantibodies in celiac disease target clustered, N-terminal epitopes not displayed on the surface of cells 1
    Article Snippet: Mouse TG2 was obtained by extraction of RNA from mouse liver biopsies and cloning of TG2-encoding cDNA into the pET-28a vector (Novagen) as described for human TG2 ( ). .. Mutations were introduced in the TG2 sequence using the QuickChange Site-directed Mutagenesis Kit (Stratagene) and the mutations were confirmed by sequencing (GATC Biotech).

    Article Title: Mechanism of protective immunity by vaccination with recombinant Echinococcus granulosus glutathione S-transferase (Chinese strain) in mice
    Article Snippet: The target fragment was purified using a gel cleanup kit (SBS Genetech, Co., Ltd.) and inserted between the Eco RI and Not I sites of the expression vector pET-28a (Novagen; Merck KGaA, Darmstadt, Germany). .. The recombinant vector EgGST/pET-28a was identified by restriction digestion using Eco RI and Not I and the EgGST insert was verified by sequencing, performed by Sangon Biotech Co., Ltd. (Shanghai, China).

    Article Title: Crystallization and preliminary X-ray crystallographic studies of fatty acid-CoA racemase from Mycobacterium tuberculosis H37Rv
    Article Snippet: The PCR product was purified, digested with Nde I and Xho I and ligated into the pET-28a vector (Novagen). .. After confirmation of the DNA sequence, the resulting vector was introduced into the Escherichia coli BL21 (DE3) strain grown at 310 K in LB medium with kanamycin (50 µg ml−1 ).

    Article Title: Detection of anti-preS1 antibodies for recovery of hepatitis B patients by immunoassay
    Article Snippet: Expression vector pET-28a (+) was purchased from Novagen. .. DNA sequence kit was from USB.

    Sonication:

    Article Title: Differential regulation of dopaminergic gene expression by Er81
    Article Snippet: ER81 used in the EMSA experiments was generated by over-expression in recombinant pLysS E. coli bacteria containing a pET-28A expression plasmid (Novagen) containing an Er81 cDNA isolated from mouse OB RNA using the Oligotex mRNA mini-kit (Qiagen). .. Resuspended cells were then sonicated, and cellular debris removed via centrifugation.

    Article Title: Bmserpin2 Is Involved in BmNPV Infection by Suppressing Melanization in Bombyx mori
    Article Snippet: Recombinant Bmserpin2 and Antibody Preparation The cloned cDNA fragment that contained mature Bmserpin2 ligated into the expression vector pET-28a (Novagen) with restriction enzyme sites BamH1 and Xho1 , and the insertion was confirmed by DNA sequencing in the recombinant BmSerpin2-pET-28a plasmid and then the recombinant plasmid was transformed into Escherichia coli BL21 (Novagen) competent cells which were grown in fresh Luria-Bertani (LB) medium. .. After centrifugation at 7500× g for 10 min at 4 °C, bacteria were collected and sonicated in phosphate-buffered saline (PBS).

    Recombinant:

    Article Title: Molecular Characterization and Identification of Target Protein of an Important Vesicle Trafficking Gene AlRab7 from a Salt Excreting Halophyte Aeluropus lagopoides
    Article Snippet: Paragraph title: Cloning of AlRab7 into an Escherichia coli expression vector and purification of the recombinant protein ... The amplified product was gel purified, digested, and cloned in pET 28a expression vector (Novagen).

    Article Title: Transglutaminase 2-specific autoantibodies in celiac disease target clustered, N-terminal epitopes not displayed on the surface of cells 1
    Article Snippet: His-tagged recombinant human TG2 was either expressed in E. coli and purified as previously described ( ) or obtained from Phadia as purified protein expressed in Sf9 insect cells. .. Mouse TG2 was obtained by extraction of RNA from mouse liver biopsies and cloning of TG2-encoding cDNA into the pET-28a vector (Novagen) as described for human TG2 ( ).

    Article Title: A broad pH range and processive chitinase from a metagenome library
    Article Snippet: .. E.coli BL21(DE3) and vector pET-28a(+) (Novagen, USA) were used as the recombinant protein expression system. .. Chemicals and enzymes FideliTaq PCR Master Mix (USB, USA) was used for DNA amplification.

    Article Title: Mechanism of protective immunity by vaccination with recombinant Echinococcus granulosus glutathione S-transferase (Chinese strain) in mice
    Article Snippet: The target fragment was purified using a gel cleanup kit (SBS Genetech, Co., Ltd.) and inserted between the Eco RI and Not I sites of the expression vector pET-28a (Novagen; Merck KGaA, Darmstadt, Germany). .. The recombinant vector EgGST/pET-28a was identified by restriction digestion using Eco RI and Not I and the EgGST insert was verified by sequencing, performed by Sangon Biotech Co., Ltd. (Shanghai, China).

    Article Title: Crystallization and preliminary X-ray crystallographic studies of fatty acid-CoA racemase from Mycobacterium tuberculosis H37Rv
    Article Snippet: The PCR product was purified, digested with Nde I and Xho I and ligated into the pET-28a vector (Novagen). .. Expression of the recombinant fatty acid-CoA racemase fused to a His tag at the N-­terminus was induced by 0.5 m M isopropyl- d -thiogalactopyranoside at an optical density of about 0.45 at 600 nm.

    Article Title: Differential regulation of dopaminergic gene expression by Er81
    Article Snippet: .. ER81 used in the EMSA experiments was generated by over-expression in recombinant pLysS E. coli bacteria containing a pET-28A expression plasmid (Novagen) containing an Er81 cDNA isolated from mouse OB RNA using the Oligotex mRNA mini-kit (Qiagen). .. ER81 expression was induced with 1mMIPTG in bacterial cultures and harvested 6 hours after induction.

    Article Title: Bmserpin2 Is Involved in BmNPV Infection by Suppressing Melanization in Bombyx mori
    Article Snippet: .. Recombinant Bmserpin2 and Antibody Preparation The cloned cDNA fragment that contained mature Bmserpin2 ligated into the expression vector pET-28a (Novagen) with restriction enzyme sites BamH1 and Xho1 , and the insertion was confirmed by DNA sequencing in the recombinant BmSerpin2-pET-28a plasmid and then the recombinant plasmid was transformed into Escherichia coli BL21 (Novagen) competent cells which were grown in fresh Luria-Bertani (LB) medium. .. The positive plasmid was induced by isopropyl β-D-thiogalactoside (IPTG) (1 mM) to induce recombinant Bmserpin2 expression for 6 h at 37 °C.

    Article Title: Molecular Characterization, Tissue Distribution, Subcellular Localization and Actin-Sequestering Function of a Thymosin Protein from Silkworm
    Article Snippet: .. Construction of Recombinant Plasmids We used pHelix-BmTHY constructed by our laboratory as a template to amplify BmTHY ORF by PCR, with primers complementary to the flanking sequences of the ORF with Eco RI and Xho I recognition sites as follows: P1 : 5′- CGC GAATT CATGGCCTGCTCC -3′ ; P2 : 5′- CCG CTCGAG TCAAGCTGATTTCTCTT -3′ ; The PCR products were purified after electrophoresis on 1% agarose gel using the PCR Rapid Purification Kit (BioDev-Tech, China).After digestion with Eco RI and Xho I, the purified PCR products were subcloned into the expression vector pET-28a (Novagen, Darmstadt, Germany) using T4 DNA ligase (Promega, USA) and transformed into E. coli TG1 cells (maintained in our laboratory) for screening purposes. .. A positive colony with BmTHY gene in the plasmid was identified by double digestion of the plasmid, followed by analysis on 1% agarose gel electrophoresis and was subsequently verified by DNA sequencing.

    Nucleic Acid Electrophoresis:

    Article Title: Hfq, a Novel Pleiotropic Regulator of Virulence-Associated Genes in Francisella tularensis ▿
    Article Snippet: The hfq gene of LVS was cloned into the pET-28a expression vector (Novagen) to create a protein with a histidine tag at the N terminus. .. Aliquots of F. tularensis LVS (and the Δ hfq strain as a control) were harvested at different cell densities, and equal amounts of cell lysate (corresponding to 2 × 108 bacteria per well) were deposited on a 12% sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis gel.

    Mutagenesis:

    Article Title: Transglutaminase 2-specific autoantibodies in celiac disease target clustered, N-terminal epitopes not displayed on the surface of cells 1
    Article Snippet: Mouse TG2 was obtained by extraction of RNA from mouse liver biopsies and cloning of TG2-encoding cDNA into the pET-28a vector (Novagen) as described for human TG2 ( ). .. Mutations were introduced in the TG2 sequence using the QuickChange Site-directed Mutagenesis Kit (Stratagene) and the mutations were confirmed by sequencing (GATC Biotech).

    Isolation:

    Article Title: Differential regulation of dopaminergic gene expression by Er81
    Article Snippet: .. ER81 used in the EMSA experiments was generated by over-expression in recombinant pLysS E. coli bacteria containing a pET-28A expression plasmid (Novagen) containing an Er81 cDNA isolated from mouse OB RNA using the Oligotex mRNA mini-kit (Qiagen). .. ER81 expression was induced with 1mMIPTG in bacterial cultures and harvested 6 hours after induction.

    Subcloning:

    Article Title: A broad pH range and processive chitinase from a metagenome library
    Article Snippet: E. coli DH10B and the vectors pUC18 and pCR2.1 (Invitrogen Life Technologies, USA) were used for subcloning steps. .. E.coli BL21(DE3) and vector pET-28a(+) (Novagen, USA) were used as the recombinant protein expression system.

    Article Title: Mechanism of protective immunity by vaccination with recombinant Echinococcus granulosus glutathione S-transferase (Chinese strain) in mice
    Article Snippet: Paragraph title: Subcloning of EgGST gene into expression plasmid vector ... The target fragment was purified using a gel cleanup kit (SBS Genetech, Co., Ltd.) and inserted between the Eco RI and Not I sites of the expression vector pET-28a (Novagen; Merck KGaA, Darmstadt, Germany).

    Electrophoretic Mobility Shift Assay:

    Article Title: Differential regulation of dopaminergic gene expression by Er81
    Article Snippet: Paragraph title: Electromobility gel shift assays (EMSA) ... ER81 used in the EMSA experiments was generated by over-expression in recombinant pLysS E. coli bacteria containing a pET-28A expression plasmid (Novagen) containing an Er81 cDNA isolated from mouse OB RNA using the Oligotex mRNA mini-kit (Qiagen).

    Purification:

    Article Title: Molecular Characterization and Identification of Target Protein of an Important Vesicle Trafficking Gene AlRab7 from a Salt Excreting Halophyte Aeluropus lagopoides
    Article Snippet: .. The amplified product was gel purified, digested, and cloned in pET 28a expression vector (Novagen). .. The recombinant plasmid was transformed in BL21 (DE3) star Escherichia coli strain and grown in Luria Bertani (LB) medium at 37°C to OD600 0.5.

    Article Title: Transglutaminase 2-specific autoantibodies in celiac disease target clustered, N-terminal epitopes not displayed on the surface of cells 1
    Article Snippet: His-tagged recombinant human TG2 was either expressed in E. coli and purified as previously described ( ) or obtained from Phadia as purified protein expressed in Sf9 insect cells. .. Mouse TG2 was obtained by extraction of RNA from mouse liver biopsies and cloning of TG2-encoding cDNA into the pET-28a vector (Novagen) as described for human TG2 ( ).

    Article Title: Mechanism of protective immunity by vaccination with recombinant Echinococcus granulosus glutathione S-transferase (Chinese strain) in mice
    Article Snippet: .. The target fragment was purified using a gel cleanup kit (SBS Genetech, Co., Ltd.) and inserted between the Eco RI and Not I sites of the expression vector pET-28a (Novagen; Merck KGaA, Darmstadt, Germany). .. The recombinant vector EgGST/pET-28a was identified by restriction digestion using Eco RI and Not I and the EgGST insert was verified by sequencing, performed by Sangon Biotech Co., Ltd. (Shanghai, China).

    Article Title: Hfq, a Novel Pleiotropic Regulator of Virulence-Associated Genes in Francisella tularensis ▿
    Article Snippet: The hfq gene of LVS was cloned into the pET-28a expression vector (Novagen) to create a protein with a histidine tag at the N terminus. .. Antiserum was purified by incubation with an F. tularensis Δ hfq bacterial lysate before it was used for immunoblotting.

    Article Title: Crystallization and preliminary X-ray crystallographic studies of fatty acid-CoA racemase from Mycobacterium tuberculosis H37Rv
    Article Snippet: .. The PCR product was purified, digested with Nde I and Xho I and ligated into the pET-28a vector (Novagen). .. After confirmation of the DNA sequence, the resulting vector was introduced into the Escherichia coli BL21 (DE3) strain grown at 310 K in LB medium with kanamycin (50 µg ml−1 ).

    Article Title: Molecular Characterization, Tissue Distribution, Subcellular Localization and Actin-Sequestering Function of a Thymosin Protein from Silkworm
    Article Snippet: .. Construction of Recombinant Plasmids We used pHelix-BmTHY constructed by our laboratory as a template to amplify BmTHY ORF by PCR, with primers complementary to the flanking sequences of the ORF with Eco RI and Xho I recognition sites as follows: P1 : 5′- CGC GAATT CATGGCCTGCTCC -3′ ; P2 : 5′- CCG CTCGAG TCAAGCTGATTTCTCTT -3′ ; The PCR products were purified after electrophoresis on 1% agarose gel using the PCR Rapid Purification Kit (BioDev-Tech, China).After digestion with Eco RI and Xho I, the purified PCR products were subcloned into the expression vector pET-28a (Novagen, Darmstadt, Germany) using T4 DNA ligase (Promega, USA) and transformed into E. coli TG1 cells (maintained in our laboratory) for screening purposes. .. A positive colony with BmTHY gene in the plasmid was identified by double digestion of the plasmid, followed by analysis on 1% agarose gel electrophoresis and was subsequently verified by DNA sequencing.

    Polymerase Chain Reaction:

    Article Title: Molecular Characterization and Identification of Target Protein of an Important Vesicle Trafficking Gene AlRab7 from a Salt Excreting Halophyte Aeluropus lagopoides
    Article Snippet: Using these primers the complete coding sequence for the AlRab7 was PCR amplified (150 ng of each primer, 200 μM dNTPs, 2.5 U Taq DNA polymerase in a 50 μL reaction, with 94°C, 5 min for 1 cycle, 94°C, 1 min, 55°C, 1 min, and 72°C, 1 min for 30 cycles; and 72°C, 7 min for 1 cycle) using Aeluropus cDNA clone as template. .. The amplified product was gel purified, digested, and cloned in pET 28a expression vector (Novagen).

    Article Title: QnrS1 structure–activity relationships
    Article Snippet: A DNA fragment encompassing the qnrS1 gene was amplified by PCR from plasmid pMG306 (GenBank accession no. ) using the primers listed in Table . .. The resulting DNA fragment was double-digested with SacI and XhoI (New England Biolabs, Ipswich, MA, USA) and cloned into expression vector pET-28a (Novagen, USA).

    Article Title: Crystallization and preliminary X-ray crystallographic studies of fatty acid-CoA racemase from Mycobacterium tuberculosis H37Rv
    Article Snippet: .. The PCR product was purified, digested with Nde I and Xho I and ligated into the pET-28a vector (Novagen). .. After confirmation of the DNA sequence, the resulting vector was introduced into the Escherichia coli BL21 (DE3) strain grown at 310 K in LB medium with kanamycin (50 µg ml−1 ).

    Article Title: Molecular Characterization, Tissue Distribution, Subcellular Localization and Actin-Sequestering Function of a Thymosin Protein from Silkworm
    Article Snippet: .. Construction of Recombinant Plasmids We used pHelix-BmTHY constructed by our laboratory as a template to amplify BmTHY ORF by PCR, with primers complementary to the flanking sequences of the ORF with Eco RI and Xho I recognition sites as follows: P1 : 5′- CGC GAATT CATGGCCTGCTCC -3′ ; P2 : 5′- CCG CTCGAG TCAAGCTGATTTCTCTT -3′ ; The PCR products were purified after electrophoresis on 1% agarose gel using the PCR Rapid Purification Kit (BioDev-Tech, China).After digestion with Eco RI and Xho I, the purified PCR products were subcloned into the expression vector pET-28a (Novagen, Darmstadt, Germany) using T4 DNA ligase (Promega, USA) and transformed into E. coli TG1 cells (maintained in our laboratory) for screening purposes. .. A positive colony with BmTHY gene in the plasmid was identified by double digestion of the plasmid, followed by analysis on 1% agarose gel electrophoresis and was subsequently verified by DNA sequencing.

    Plasmid Preparation:

    Article Title: Molecular Characterization and Identification of Target Protein of an Important Vesicle Trafficking Gene AlRab7 from a Salt Excreting Halophyte Aeluropus lagopoides
    Article Snippet: .. The amplified product was gel purified, digested, and cloned in pET 28a expression vector (Novagen). .. The recombinant plasmid was transformed in BL21 (DE3) star Escherichia coli strain and grown in Luria Bertani (LB) medium at 37°C to OD600 0.5.

    Article Title: Tailoring the properties of (catalytically)-active inclusion bodies
    Article Snippet: .. If not stated otherwise, all gene fusions consisted of gene fragments coding for a coiled-coil domain (here TDoT or 3HAMP), a linker polypeptide, consisting of a protease Factor Xa cleavage site and a triple (GGGS)3 and the respective target proteins/enzymes cloned into a pET-28a vector (Novagen, Merck KGaA, Frankfurt, Germany). ..

    Article Title: Transglutaminase 2-specific autoantibodies in celiac disease target clustered, N-terminal epitopes not displayed on the surface of cells 1
    Article Snippet: .. Mouse TG2 was obtained by extraction of RNA from mouse liver biopsies and cloning of TG2-encoding cDNA into the pET-28a vector (Novagen) as described for human TG2 ( ). .. Mouse TG2 and human TG2 mutants were produced in E. coli in the same way as wild-type human TG2.

    Article Title: 1H, 13C, and 15N resonance assignments and secondary structure prediction of the full-length transition state regulator AbrB from Bacillus anthracis
    Article Snippet: .. Full length AbrB (residues 1–94) from B. anthracis was cloned into expression vector pET-28a (Novagen) with a Thrombin cleavable N-terminal histidine tag and transformed into BL21(DE3) cells (Genesee) for expression. ..

    Article Title: A broad pH range and processive chitinase from a metagenome library
    Article Snippet: .. E.coli BL21(DE3) and vector pET-28a(+) (Novagen, USA) were used as the recombinant protein expression system. .. Chemicals and enzymes FideliTaq PCR Master Mix (USB, USA) was used for DNA amplification.

    Article Title: Mechanism of protective immunity by vaccination with recombinant Echinococcus granulosus glutathione S-transferase (Chinese strain) in mice
    Article Snippet: .. The target fragment was purified using a gel cleanup kit (SBS Genetech, Co., Ltd.) and inserted between the Eco RI and Not I sites of the expression vector pET-28a (Novagen; Merck KGaA, Darmstadt, Germany). .. The recombinant vector EgGST/pET-28a was identified by restriction digestion using Eco RI and Not I and the EgGST insert was verified by sequencing, performed by Sangon Biotech Co., Ltd. (Shanghai, China).

    Article Title: Hfq, a Novel Pleiotropic Regulator of Virulence-Associated Genes in Francisella tularensis ▿
    Article Snippet: .. The hfq gene of LVS was cloned into the pET-28a expression vector (Novagen) to create a protein with a histidine tag at the N terminus. ..

    Article Title: High Level Expression and Purification of the Clinically Active Antimicrobial Peptide P-113 in Escherichia coli
    Article Snippet: .. The expression vector pET-28a and host BL21 (DE3) were purchased from Novagen (Merck Millipore, Burlington, MA, USA). .. Restriction enzymes and T4 ligase were purchased from Thermo Scientific (Waltham, MA, USA).

    Article Title: QnrS1 structure–activity relationships
    Article Snippet: .. The resulting DNA fragment was double-digested with SacI and XhoI (New England Biolabs, Ipswich, MA, USA) and cloned into expression vector pET-28a (Novagen, USA). .. Plasmid pET-28a:QnrS1 was transformed into E. coli BL21(DE3) (New England Biolabs) and plated on Luria–Bertani (LB) agar (Becton Dickinson, USA) containing 100 mg/L kanamycin to maintain the plasmid.

    Article Title: Crystallization and preliminary X-ray crystallographic studies of fatty acid-CoA racemase from Mycobacterium tuberculosis H37Rv
    Article Snippet: .. The PCR product was purified, digested with Nde I and Xho I and ligated into the pET-28a vector (Novagen). .. After confirmation of the DNA sequence, the resulting vector was introduced into the Escherichia coli BL21 (DE3) strain grown at 310 K in LB medium with kanamycin (50 µg ml−1 ).

    Article Title: Differential regulation of dopaminergic gene expression by Er81
    Article Snippet: .. ER81 used in the EMSA experiments was generated by over-expression in recombinant pLysS E. coli bacteria containing a pET-28A expression plasmid (Novagen) containing an Er81 cDNA isolated from mouse OB RNA using the Oligotex mRNA mini-kit (Qiagen). .. ER81 expression was induced with 1mMIPTG in bacterial cultures and harvested 6 hours after induction.

    Article Title: High Level Expression and Purification of the Clinically Active Antimicrobial Peptide P-113 in Escherichia coli
    Article Snippet: .. Materials and Methods The expression vector pET-28a and host BL21 (DE3) were purchased from Novagen (Merck Millipore, Burlington, MA, USA). .. Restriction enzymes and T4 ligase were purchased from Thermo Scientific (Waltham, MA, USA).

    Article Title: Bmserpin2 Is Involved in BmNPV Infection by Suppressing Melanization in Bombyx mori
    Article Snippet: .. Recombinant Bmserpin2 and Antibody Preparation The cloned cDNA fragment that contained mature Bmserpin2 ligated into the expression vector pET-28a (Novagen) with restriction enzyme sites BamH1 and Xho1 , and the insertion was confirmed by DNA sequencing in the recombinant BmSerpin2-pET-28a plasmid and then the recombinant plasmid was transformed into Escherichia coli BL21 (Novagen) competent cells which were grown in fresh Luria-Bertani (LB) medium. .. The positive plasmid was induced by isopropyl β-D-thiogalactoside (IPTG) (1 mM) to induce recombinant Bmserpin2 expression for 6 h at 37 °C.

    Article Title: Molecular Characterization, Tissue Distribution, Subcellular Localization and Actin-Sequestering Function of a Thymosin Protein from Silkworm
    Article Snippet: .. Construction of Recombinant Plasmids We used pHelix-BmTHY constructed by our laboratory as a template to amplify BmTHY ORF by PCR, with primers complementary to the flanking sequences of the ORF with Eco RI and Xho I recognition sites as follows: P1 : 5′- CGC GAATT CATGGCCTGCTCC -3′ ; P2 : 5′- CCG CTCGAG TCAAGCTGATTTCTCTT -3′ ; The PCR products were purified after electrophoresis on 1% agarose gel using the PCR Rapid Purification Kit (BioDev-Tech, China).After digestion with Eco RI and Xho I, the purified PCR products were subcloned into the expression vector pET-28a (Novagen, Darmstadt, Germany) using T4 DNA ligase (Promega, USA) and transformed into E. coli TG1 cells (maintained in our laboratory) for screening purposes. .. A positive colony with BmTHY gene in the plasmid was identified by double digestion of the plasmid, followed by analysis on 1% agarose gel electrophoresis and was subsequently verified by DNA sequencing.

    Article Title: Detection of anti-preS1 antibodies for recovery of hepatitis B patients by immunoassay
    Article Snippet: .. Expression vector pET-28a (+) was purchased from Novagen. ..

    Positron Emission Tomography:

    Article Title: Molecular Characterization and Identification of Target Protein of an Important Vesicle Trafficking Gene AlRab7 from a Salt Excreting Halophyte Aeluropus lagopoides
    Article Snippet: .. The amplified product was gel purified, digested, and cloned in pET 28a expression vector (Novagen). .. The recombinant plasmid was transformed in BL21 (DE3) star Escherichia coli strain and grown in Luria Bertani (LB) medium at 37°C to OD600 0.5.

    Article Title: Tailoring the properties of (catalytically)-active inclusion bodies
    Article Snippet: .. If not stated otherwise, all gene fusions consisted of gene fragments coding for a coiled-coil domain (here TDoT or 3HAMP), a linker polypeptide, consisting of a protease Factor Xa cleavage site and a triple (GGGS)3 and the respective target proteins/enzymes cloned into a pET-28a vector (Novagen, Merck KGaA, Frankfurt, Germany). ..

    Article Title: Transglutaminase 2-specific autoantibodies in celiac disease target clustered, N-terminal epitopes not displayed on the surface of cells 1
    Article Snippet: .. Mouse TG2 was obtained by extraction of RNA from mouse liver biopsies and cloning of TG2-encoding cDNA into the pET-28a vector (Novagen) as described for human TG2 ( ). .. Mouse TG2 and human TG2 mutants were produced in E. coli in the same way as wild-type human TG2.

    Article Title: 1H, 13C, and 15N resonance assignments and secondary structure prediction of the full-length transition state regulator AbrB from Bacillus anthracis
    Article Snippet: .. Full length AbrB (residues 1–94) from B. anthracis was cloned into expression vector pET-28a (Novagen) with a Thrombin cleavable N-terminal histidine tag and transformed into BL21(DE3) cells (Genesee) for expression. ..

    Article Title: A broad pH range and processive chitinase from a metagenome library
    Article Snippet: .. E.coli BL21(DE3) and vector pET-28a(+) (Novagen, USA) were used as the recombinant protein expression system. .. Chemicals and enzymes FideliTaq PCR Master Mix (USB, USA) was used for DNA amplification.

    Article Title: Mechanism of protective immunity by vaccination with recombinant Echinococcus granulosus glutathione S-transferase (Chinese strain) in mice
    Article Snippet: .. The target fragment was purified using a gel cleanup kit (SBS Genetech, Co., Ltd.) and inserted between the Eco RI and Not I sites of the expression vector pET-28a (Novagen; Merck KGaA, Darmstadt, Germany). .. The recombinant vector EgGST/pET-28a was identified by restriction digestion using Eco RI and Not I and the EgGST insert was verified by sequencing, performed by Sangon Biotech Co., Ltd. (Shanghai, China).

    Article Title: Hfq, a Novel Pleiotropic Regulator of Virulence-Associated Genes in Francisella tularensis ▿
    Article Snippet: .. The hfq gene of LVS was cloned into the pET-28a expression vector (Novagen) to create a protein with a histidine tag at the N terminus. ..

    Article Title: High Level Expression and Purification of the Clinically Active Antimicrobial Peptide P-113 in Escherichia coli
    Article Snippet: .. The expression vector pET-28a and host BL21 (DE3) were purchased from Novagen (Merck Millipore, Burlington, MA, USA). .. Restriction enzymes and T4 ligase were purchased from Thermo Scientific (Waltham, MA, USA).

    Article Title: QnrS1 structure–activity relationships
    Article Snippet: .. The resulting DNA fragment was double-digested with SacI and XhoI (New England Biolabs, Ipswich, MA, USA) and cloned into expression vector pET-28a (Novagen, USA). .. Plasmid pET-28a:QnrS1 was transformed into E. coli BL21(DE3) (New England Biolabs) and plated on Luria–Bertani (LB) agar (Becton Dickinson, USA) containing 100 mg/L kanamycin to maintain the plasmid.

    Article Title: Crystallization and preliminary X-ray crystallographic studies of fatty acid-CoA racemase from Mycobacterium tuberculosis H37Rv
    Article Snippet: .. The PCR product was purified, digested with Nde I and Xho I and ligated into the pET-28a vector (Novagen). .. After confirmation of the DNA sequence, the resulting vector was introduced into the Escherichia coli BL21 (DE3) strain grown at 310 K in LB medium with kanamycin (50 µg ml−1 ).

    Article Title: Differential regulation of dopaminergic gene expression by Er81
    Article Snippet: .. ER81 used in the EMSA experiments was generated by over-expression in recombinant pLysS E. coli bacteria containing a pET-28A expression plasmid (Novagen) containing an Er81 cDNA isolated from mouse OB RNA using the Oligotex mRNA mini-kit (Qiagen). .. ER81 expression was induced with 1mMIPTG in bacterial cultures and harvested 6 hours after induction.

    Article Title: High Level Expression and Purification of the Clinically Active Antimicrobial Peptide P-113 in Escherichia coli
    Article Snippet: .. Materials and Methods The expression vector pET-28a and host BL21 (DE3) were purchased from Novagen (Merck Millipore, Burlington, MA, USA). .. Restriction enzymes and T4 ligase were purchased from Thermo Scientific (Waltham, MA, USA).

    Article Title: Bmserpin2 Is Involved in BmNPV Infection by Suppressing Melanization in Bombyx mori
    Article Snippet: .. Recombinant Bmserpin2 and Antibody Preparation The cloned cDNA fragment that contained mature Bmserpin2 ligated into the expression vector pET-28a (Novagen) with restriction enzyme sites BamH1 and Xho1 , and the insertion was confirmed by DNA sequencing in the recombinant BmSerpin2-pET-28a plasmid and then the recombinant plasmid was transformed into Escherichia coli BL21 (Novagen) competent cells which were grown in fresh Luria-Bertani (LB) medium. .. The positive plasmid was induced by isopropyl β-D-thiogalactoside (IPTG) (1 mM) to induce recombinant Bmserpin2 expression for 6 h at 37 °C.

    Article Title: Molecular Characterization, Tissue Distribution, Subcellular Localization and Actin-Sequestering Function of a Thymosin Protein from Silkworm
    Article Snippet: .. Construction of Recombinant Plasmids We used pHelix-BmTHY constructed by our laboratory as a template to amplify BmTHY ORF by PCR, with primers complementary to the flanking sequences of the ORF with Eco RI and Xho I recognition sites as follows: P1 : 5′- CGC GAATT CATGGCCTGCTCC -3′ ; P2 : 5′- CCG CTCGAG TCAAGCTGATTTCTCTT -3′ ; The PCR products were purified after electrophoresis on 1% agarose gel using the PCR Rapid Purification Kit (BioDev-Tech, China).After digestion with Eco RI and Xho I, the purified PCR products were subcloned into the expression vector pET-28a (Novagen, Darmstadt, Germany) using T4 DNA ligase (Promega, USA) and transformed into E. coli TG1 cells (maintained in our laboratory) for screening purposes. .. A positive colony with BmTHY gene in the plasmid was identified by double digestion of the plasmid, followed by analysis on 1% agarose gel electrophoresis and was subsequently verified by DNA sequencing.

    Article Title: Detection of anti-preS1 antibodies for recovery of hepatitis B patients by immunoassay
    Article Snippet: .. Expression vector pET-28a (+) was purchased from Novagen. ..

    Agarose Gel Electrophoresis:

    Article Title: Molecular Characterization, Tissue Distribution, Subcellular Localization and Actin-Sequestering Function of a Thymosin Protein from Silkworm
    Article Snippet: .. Construction of Recombinant Plasmids We used pHelix-BmTHY constructed by our laboratory as a template to amplify BmTHY ORF by PCR, with primers complementary to the flanking sequences of the ORF with Eco RI and Xho I recognition sites as follows: P1 : 5′- CGC GAATT CATGGCCTGCTCC -3′ ; P2 : 5′- CCG CTCGAG TCAAGCTGATTTCTCTT -3′ ; The PCR products were purified after electrophoresis on 1% agarose gel using the PCR Rapid Purification Kit (BioDev-Tech, China).After digestion with Eco RI and Xho I, the purified PCR products were subcloned into the expression vector pET-28a (Novagen, Darmstadt, Germany) using T4 DNA ligase (Promega, USA) and transformed into E. coli TG1 cells (maintained in our laboratory) for screening purposes. .. A positive colony with BmTHY gene in the plasmid was identified by double digestion of the plasmid, followed by analysis on 1% agarose gel electrophoresis and was subsequently verified by DNA sequencing.

    Produced:

    Article Title: Transglutaminase 2-specific autoantibodies in celiac disease target clustered, N-terminal epitopes not displayed on the surface of cells 1
    Article Snippet: Mouse TG2 was obtained by extraction of RNA from mouse liver biopsies and cloning of TG2-encoding cDNA into the pET-28a vector (Novagen) as described for human TG2 ( ). .. Mouse TG2 and human TG2 mutants were produced in E. coli in the same way as wild-type human TG2.

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  • 99
    Millipore plasmid pet 28a
    Validation of nucleic acid digestion by pepsin. a , Digestion of various DNA and RNA by commercial porcine pepsin. Lane 1, 2: salmon sperm DNA; Lane 3, 4: λ DNA; Lane 5, 6: <t>pET-28a;</t> Lane 7, 8: M13mp18; Lane 9, 10: RNA ladder. Other conditions: 4.0 mg ml −1 of pepsin, NaH 2 PO 4 buffer (25 mM, pH 3.8, including 200 mM NaCl), 37 °C, 5 h. For RNA, the digestion time was 1 h. b , Effect of alkaline conditions on pepsin NA digestion. Lane 1, original λ DNA; Lane 2, digested by active pepsin; Lane 3, digested by NaOH-pretreated pepsin. c, Digestion of λ DNA by commercial porcine pepsin (Lane P), recombinant pepsin (Lane rP) and mutant pepsin (Lane mP). Conditions: 0.15 mg ml −1 enzymes, pH 3.8, 37 °C, 12 h. A 0.8% agarose gel was used for electrophoresis.
    Plasmid Pet 28a, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 375 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/plasmid pet 28a/product/Millipore
    Average 99 stars, based on 375 article reviews
    Price from $9.99 to $1999.99
    plasmid pet 28a - by Bioz Stars, 2020-02
    99/100 stars
      Buy from Supplier

    99
    Millipore pet 28a
    Validation of nucleic acid digestion by pepsin. a , Digestion of various DNA and RNA by commercial porcine pepsin. Lane 1, 2: salmon sperm DNA; Lane 3, 4: λ DNA; Lane 5, 6: <t>pET-28a;</t> Lane 7, 8: M13mp18; Lane 9, 10: RNA ladder. Other conditions: 4.0 mg ml −1 of pepsin, NaH 2 PO 4 buffer (25 mM, pH 3.8, including 200 mM NaCl), 37 °C, 5 h. For RNA, the digestion time was 1 h. b , Effect of alkaline conditions on pepsin NA digestion. Lane 1, original λ DNA; Lane 2, digested by active pepsin; Lane 3, digested by NaOH-pretreated pepsin. c, Digestion of λ DNA by commercial porcine pepsin (Lane P), recombinant pepsin (Lane rP) and mutant pepsin (Lane mP). Conditions: 0.15 mg ml −1 enzymes, pH 3.8, 37 °C, 12 h. A 0.8% agarose gel was used for electrophoresis.
    Pet 28a, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 452 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pet 28a/product/Millipore
    Average 99 stars, based on 452 article reviews
    Price from $9.99 to $1999.99
    pet 28a - by Bioz Stars, 2020-02
    99/100 stars
      Buy from Supplier

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    Validation of nucleic acid digestion by pepsin. a , Digestion of various DNA and RNA by commercial porcine pepsin. Lane 1, 2: salmon sperm DNA; Lane 3, 4: λ DNA; Lane 5, 6: pET-28a; Lane 7, 8: M13mp18; Lane 9, 10: RNA ladder. Other conditions: 4.0 mg ml −1 of pepsin, NaH 2 PO 4 buffer (25 mM, pH 3.8, including 200 mM NaCl), 37 °C, 5 h. For RNA, the digestion time was 1 h. b , Effect of alkaline conditions on pepsin NA digestion. Lane 1, original λ DNA; Lane 2, digested by active pepsin; Lane 3, digested by NaOH-pretreated pepsin. c, Digestion of λ DNA by commercial porcine pepsin (Lane P), recombinant pepsin (Lane rP) and mutant pepsin (Lane mP). Conditions: 0.15 mg ml −1 enzymes, pH 3.8, 37 °C, 12 h. A 0.8% agarose gel was used for electrophoresis.

    Journal: Scientific Reports

    Article Title: Digestion of Nucleic Acids Starts in the Stomach

    doi: 10.1038/srep11936

    Figure Lengend Snippet: Validation of nucleic acid digestion by pepsin. a , Digestion of various DNA and RNA by commercial porcine pepsin. Lane 1, 2: salmon sperm DNA; Lane 3, 4: λ DNA; Lane 5, 6: pET-28a; Lane 7, 8: M13mp18; Lane 9, 10: RNA ladder. Other conditions: 4.0 mg ml −1 of pepsin, NaH 2 PO 4 buffer (25 mM, pH 3.8, including 200 mM NaCl), 37 °C, 5 h. For RNA, the digestion time was 1 h. b , Effect of alkaline conditions on pepsin NA digestion. Lane 1, original λ DNA; Lane 2, digested by active pepsin; Lane 3, digested by NaOH-pretreated pepsin. c, Digestion of λ DNA by commercial porcine pepsin (Lane P), recombinant pepsin (Lane rP) and mutant pepsin (Lane mP). Conditions: 0.15 mg ml −1 enzymes, pH 3.8, 37 °C, 12 h. A 0.8% agarose gel was used for electrophoresis.

    Article Snippet: Plasmid pET-28a (5.4 kb) was purchased from EMD Chemicals Inc. (Novagen, D00131614) with concentration of 500 μg ml−1 , and was diluted into 210 μg ml−1 before use.

    Techniques: Positron Emission Tomography, Recombinant, Mutagenesis, Agarose Gel Electrophoresis, Electrophoresis