bl21 de3 plyss  (Millipore)


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    Structured Review

    Millipore bl21 de3 plyss
    Bl21 De3 Plyss, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bl21 de3 plyss/product/Millipore
    Average 99 stars, based on 36 article reviews
    Price from $9.99 to $1999.99
    bl21 de3 plyss - by Bioz Stars, 2020-03
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Immunization with Recombinant Transferrin Binding Protein B Enhances Clearance of Nontypeable Haemophilus influenzae from the Rat Lung
    Article Snippet: Paragraph title: Cloning and purification of recombinant NTHI TbpB. ... To purify rTbpB, cultures of BL21 [F− ompT hsd SB (rB − mB − ) gal dcm ] (Novagen, Madison, Wis.) containing pCU17 were grown in Luria-Bertani broth supplemented with 3.6 g of glucose per liter and 100 μg of ampicillin per ml.

    Article Title: YeeV is an Escherichia coli Toxin that Inhibits Cell Division by Targeting the Cytoskeleton Proteins, FtsZ and MreB
    Article Snippet: E. coli BW25113 ( ΔaraBD ) (Datsenko and Wanner, 2000), BL21 (DE3) were grown in M9 medium at 37° C. Plasmids pET-28a- yeeV , pET-28a- ftsZ , pET-28a- mreB and pET-28a -sulA were constructed in pET-28a (Novagen) with an N-terminal His6 tag. .. To generate the N-terminal GFP-fusion plasmids, a DNA fragment of gfp from plasmid pAcGFP1 (Clontech laboratories, Inc.) was cloned into pBAD33 to generate pBAD33- gfp.

    Article Title: Construction of a Novel Liver-Targeting Fusion Interferon by Incorporation of a Plasmodium Region I-Plus Peptide
    Article Snippet: .. Bacterial Strains, Plasmids, and Culture Media E. coli strain DH5α (Novagen, USA) and BL21 (DE3; Novagen, USA) were used as the host for gene manipulation and expression of fusion protein, respectively. pMD20-T (Takara, Japan) and pET-21b (Novagen, USA) were applied for gene cloning and expression, respectively. ..

    Article Title: Molecular basis for transfer RNA recognition by the double-stranded RNA-binding domain of human dihydrouridine synthase 2
    Article Snippet: .. Protein purification hDus2 or its dsRBD (T339-K451 construct) containing both the NTE and CTE was cloned in a pET11d vector between BamHI and NcoI and expressed in BL21(DE3)pLysS or BL21(DE3) respectively (Novagen) in LB medium and was purified as previously described ( ). .. Point mutations were carried out with Q5® Site-directed mutagenesis kit (New England BioLabs) following the recommended procedure and done on dsRDB-pET11d vector.

    Centrifugation:

    Article Title: Preparation of Biologically Active Single-Chain Variable Antibody Fragments that Target the HIV-1 gp120 V3 Loop
    Article Snippet: To optimize scFv expression in various E. coli expression strains, including BL21 (DE3), Rosetta 2 (DE3) (Novagen), BL21 Gold (DE3) pLysS (Stratagene), and BL21 Star (DE3) (Invitrogen), a similar protocol is used with modifications of the growing culture (in LB with the appropriate antibiotics) at 37 °C and inducing expression with 0.5 mM IPTG. .. 10 μg/ml DNAse and 20 mM MgSO4 were added to cell suspension and incubated on ice for 30 min before centrifugation at 13,000 × g for 20 min at 4 °C.

    Amplification:

    Article Title: Zinc Metalloproteinase ProA Directly Activates Legionella pneumophila PlaC Glycerophospholipid:cholesterol Acyltransferase *
    Article Snippet: E. coli DH5α or BL21 was used for the propagation of recombinant plasmid DNA with backbones of the following vectors: pMMB2002 , pet28a(+) (Novagen), pBADTOPO (Invitrogen), pBCKS (Stratagene), pGEX-6P-1 (GE Healthcare), pGEM-T Easy (Promega GmbH), and pET160 (Invitrogen). .. Genomic and plasmid DNA were prepared, amplified, and sequenced according to standard protocols.

    Article Title: Host cell and expression engineering for development of an E. coli ketoreductase catalyst: Enhancement of formate dehydrogenase activity for regeneration of NADH
    Article Snippet: Strains and plasmids The E. coli strains used were JM109 from Promega (Madison, WI, USA), BL21 (DE3) and Rosetta 2 (DE3) from Novagen (VWR International GmbH, Vienna, Austria) and BL21 star (DE3) from Invitrogen (Carlsbad, California, USA). .. The Cb FDH gene was amplified from pETDuet_XR_FDH by a PCR using Pfu DNA polymerase and primers providing Pag I (compatible ends to Nco I) and Avr II restriction sites.

    Construct:

    Article Title: Production of bioactive liver-targeting interferon Mu-IFN-CSP by soluble prokaryotic expression
    Article Snippet: .. E. coli strain DH5α (Novagen, Madison, USA) was employed for gene manipulation and BL21 (DE3; Novagen, Madison, USA) was employed to construct recombinant expression strain. .. Recombinant IFN-CSP/pET-21b expression plasmid was obtained from Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangzhou, People’s Republic of China.

    Article Title: Soluble Expression of Humanized Anti-CD20 Single Chain Antibody in Escherichia coli by Cytoplasmic Chaperones Co-expression
    Article Snippet: Bacterial strains, media, and plasmids E. coli DH5α, BL21 (DE3) and expression vector pET22b (+) were purchased from Novagen. .. The pUC-57-huscFv construct was obtained from V. Ahmadzadeh .

    Article Title: YeeV is an Escherichia coli Toxin that Inhibits Cell Division by Targeting the Cytoskeleton Proteins, FtsZ and MreB
    Article Snippet: .. E. coli BW25113 ( ΔaraBD ) (Datsenko and Wanner, 2000), BL21 (DE3) were grown in M9 medium at 37° C. Plasmids pET-28a- yeeV , pET-28a- ftsZ , pET-28a- mreB and pET-28a -sulA were constructed in pET-28a (Novagen) with an N-terminal His6 tag. .. Plasmids pBAD33- yeeV and pBAD33- sulA were constructed from pBAD33 (Guzman et al., 1995) to tightly regulate gene expression by the addition of arabinose.

    Article Title: Soluble Expression of Humanized Anti-CD20 Single Chain Antibody in Escherichia coli by Cytoplasmic Chaperones Co-expression
    Article Snippet: E. coli DH5α, BL21 (DE3) and expression vector pET22b (+) were purchased from Novagen. .. The pUC-57-huscFv construct was obtained from V. Ahmadzadeh .

    Article Title: Molecular basis for transfer RNA recognition by the double-stranded RNA-binding domain of human dihydrouridine synthase 2
    Article Snippet: .. Protein purification hDus2 or its dsRBD (T339-K451 construct) containing both the NTE and CTE was cloned in a pET11d vector between BamHI and NcoI and expressed in BL21(DE3)pLysS or BL21(DE3) respectively (Novagen) in LB medium and was purified as previously described ( ). .. Point mutations were carried out with Q5® Site-directed mutagenesis kit (New England BioLabs) following the recommended procedure and done on dsRDB-pET11d vector.

    Nuclear Magnetic Resonance:

    Article Title: Molecular basis for transfer RNA recognition by the double-stranded RNA-binding domain of human dihydrouridine synthase 2
    Article Snippet: Protein purification hDus2 or its dsRBD (T339-K451 construct) containing both the NTE and CTE was cloned in a pET11d vector between BamHI and NcoI and expressed in BL21(DE3)pLysS or BL21(DE3) respectively (Novagen) in LB medium and was purified as previously described ( ). .. For NMR experiments, cells containing the plasmid encoding the dsRDB were grown in M9 minimal medium supplemented with 1 g/l of 15 NH4 Cl for 15 N-labeling or 1 g/l 15 NH4 Cl and 2 g/l 13 C-glucose for 13 C/15 N-labeling.

    Expressing:

    Article Title: Preparation of Biologically Active Single-Chain Variable Antibody Fragments that Target the HIV-1 gp120 V3 Loop
    Article Snippet: .. To optimize scFv expression in various E. coli expression strains, including BL21 (DE3), Rosetta 2 (DE3) (Novagen), BL21 Gold (DE3) pLysS (Stratagene), and BL21 Star (DE3) (Invitrogen), a similar protocol is used with modifications of the growing culture (in LB with the appropriate antibiotics) at 37 °C and inducing expression with 0.5 mM IPTG. .. Each frozen cell pellet was thawed on ice and resuspended in 5 ml lysis buffer containing 50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1 mM EDTA, 0.1% Triton X-100, 1 mM phenylmethylsulfonyl fluoride (PMSF), and 250 μg/ml lysozyme.

    Article Title: Bacterial Genome Partitioning: N-Terminal Domain of IncC Protein Encoded by Broad-Host-Range Plasmid RK2 Modulates Oligomerisation and DNA Binding
    Article Snippet: .. Bacterial strains and growth conditions E. coli strains used in this study were DH5α (supE44 ΔlacU61 [ϕ80lacZ ΔM15] hsdR17 recA1 endA1 gyrA96 thi-1 relA1 ) for constructing and propagating plasmids and BL21 (F− ompT , hsdSB [rB − mB − ] gal , dcm [phage DE3]) for overexpression of proteins from expression vectors with the bacteriophage T7 promoter (Novagen). ..

    Article Title: Production of bioactive liver-targeting interferon Mu-IFN-CSP by soluble prokaryotic expression
    Article Snippet: .. E. coli strain DH5α (Novagen, Madison, USA) was employed for gene manipulation and BL21 (DE3; Novagen, Madison, USA) was employed to construct recombinant expression strain. .. Recombinant IFN-CSP/pET-21b expression plasmid was obtained from Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangzhou, People’s Republic of China.

    Article Title: Soluble Expression of Humanized Anti-CD20 Single Chain Antibody in Escherichia coli by Cytoplasmic Chaperones Co-expression
    Article Snippet: .. Bacterial strains, media, and plasmids E. coli DH5α, BL21 (DE3) and expression vector pET22b (+) were purchased from Novagen. .. IPTG, Taq DNA polymerase was obtained from Promega Company.

    Article Title: YeeV is an Escherichia coli Toxin that Inhibits Cell Division by Targeting the Cytoskeleton Proteins, FtsZ and MreB
    Article Snippet: E. coli BW25113 ( ΔaraBD ) (Datsenko and Wanner, 2000), BL21 (DE3) were grown in M9 medium at 37° C. Plasmids pET-28a- yeeV , pET-28a- ftsZ , pET-28a- mreB and pET-28a -sulA were constructed in pET-28a (Novagen) with an N-terminal His6 tag. .. Plasmids pBAD33- yeeV and pBAD33- sulA were constructed from pBAD33 (Guzman et al., 1995) to tightly regulate gene expression by the addition of arabinose.

    Article Title: Soluble Expression of Humanized Anti-CD20 Single Chain Antibody in Escherichia coli by Cytoplasmic Chaperones Co-expression
    Article Snippet: .. E. coli DH5α, BL21 (DE3) and expression vector pET22b (+) were purchased from Novagen. .. IPTG, Taq DNA polymerase was obtained from Promega Company.

    Article Title: Construction of a Novel Liver-Targeting Fusion Interferon by Incorporation of a Plasmodium Region I-Plus Peptide
    Article Snippet: .. Bacterial Strains, Plasmids, and Culture Media E. coli strain DH5α (Novagen, USA) and BL21 (DE3; Novagen, USA) were used as the host for gene manipulation and expression of fusion protein, respectively. pMD20-T (Takara, Japan) and pET-21b (Novagen, USA) were applied for gene cloning and expression, respectively. ..

    Transformation Assay:

    Article Title: Preparation of Biologically Active Single-Chain Variable Antibody Fragments that Target the HIV-1 gp120 V3 Loop
    Article Snippet: A single colony of transformed cell was inoculated in 10 ml Luria-Bertani broth (LB) containing 50 μg/ml kanamycin, 34 μg/ml chloramphenicol, and 10 μg/ml tetracycline at 37 °C with shaking at 225 rpm for overnight. .. To optimize scFv expression in various E. coli expression strains, including BL21 (DE3), Rosetta 2 (DE3) (Novagen), BL21 Gold (DE3) pLysS (Stratagene), and BL21 Star (DE3) (Invitrogen), a similar protocol is used with modifications of the growing culture (in LB with the appropriate antibiotics) at 37 °C and inducing expression with 0.5 mM IPTG.

    Article Title: The pCri System: A Vector Collection for Recombinant Protein Expression and Purification
    Article Snippet: .. Chemically competent E. coli DH5α, BL21 (DE3), and Origami 2 (DE3) cells (Novagen) were prepared and transformed following Hanahan method . .. Competent cells of P. pastoris KM71H (Invitrogen) and B. subtilis WB800N (MoBiTec) were prepared according to the manufacturer's instructions.

    Over Expression:

    Article Title: Bacterial Genome Partitioning: N-Terminal Domain of IncC Protein Encoded by Broad-Host-Range Plasmid RK2 Modulates Oligomerisation and DNA Binding
    Article Snippet: .. Bacterial strains and growth conditions E. coli strains used in this study were DH5α (supE44 ΔlacU61 [ϕ80lacZ ΔM15] hsdR17 recA1 endA1 gyrA96 thi-1 relA1 ) for constructing and propagating plasmids and BL21 (F− ompT , hsdSB [rB − mB − ] gal , dcm [phage DE3]) for overexpression of proteins from expression vectors with the bacteriophage T7 promoter (Novagen). ..

    Cell Culture:

    Article Title: Reduced Renal Colonization and Enhanced Protection by Leptospiral Factor H Binding Proteins as a Multisubunit Vaccine against Leptospirosis in Hamsters
    Article Snippet: Leptospires were cultured at 30 °C in Ellinghausen–McCullough–Johnson–Harris (EMJH) medium (BD Difco™, MD, USA) supplemented with 10% bovine serum albumin (BSA) [ ]. .. Escherichia coli strains DH5α and BL21 (DE3) pLysS (Novagen, Darmstadt, Germany) were grown at 37 °C in Luria–Bertani (LB) medium with the addition of 100 µg/mL ampicillin and 30 µg/mL chloramphenicol when required.

    Polymerase Chain Reaction:

    Article Title: Immunization with Recombinant Transferrin Binding Protein B Enhances Clearance of Nontypeable Haemophilus influenzae from the Rat Lung
    Article Snippet: Agarose gel analysis of the PCR product demonstrated a ∼1.8 kb fragment, which was purified with Bresaclean resin (Bresatec Ltd. Adelaide, Australia), digested with Bam HI, and cloned into the Bam HI restriction sites in plasmid pGEX2T (Pharmacia Biotech, Uppsala, Sweden) to produce plasmid pCU17. .. To purify rTbpB, cultures of BL21 [F− ompT hsd SB (rB − mB − ) gal dcm ] (Novagen, Madison, Wis.) containing pCU17 were grown in Luria-Bertani broth supplemented with 3.6 g of glucose per liter and 100 μg of ampicillin per ml.

    Article Title: A Molecular Biological and Biochemical Investigation on Mycobacterium tuberculosis MutT Protein
    Article Snippet: Bacterial Strains, Plasmids, and Antibodies E. coli isogenic strain MK602 (leu+ mutT ) was provided by Dr. M. Sekiguchi , and BL21 (DE3) was purchased from Novagen (Madison, WI). .. Plasmids pCR®II and pET28a (+) were obtained from Invitrogen (Carlsbad, CA) and Novagen (Madison, WI), respectively.

    Article Title: The pCri System: A Vector Collection for Recombinant Protein Expression and Purification
    Article Snippet: DNA was purified from PCR reactions, enzymatic reactions, agarose gel band extractions, and vector extractions using OMEGA-Biotek purification kits. .. Chemically competent E. coli DH5α, BL21 (DE3), and Origami 2 (DE3) cells (Novagen) were prepared and transformed following Hanahan method .

    Article Title: Host cell and expression engineering for development of an E. coli ketoreductase catalyst: Enhancement of formate dehydrogenase activity for regeneration of NADH
    Article Snippet: Strains and plasmids The E. coli strains used were JM109 from Promega (Madison, WI, USA), BL21 (DE3) and Rosetta 2 (DE3) from Novagen (VWR International GmbH, Vienna, Austria) and BL21 star (DE3) from Invitrogen (Carlsbad, California, USA). .. The Cb FDH gene was amplified from pETDuet_XR_FDH by a PCR using Pfu DNA polymerase and primers providing Pag I (compatible ends to Nco I) and Avr II restriction sites.

    Recombinant:

    Article Title: Immunization with Recombinant Transferrin Binding Protein B Enhances Clearance of Nontypeable Haemophilus influenzae from the Rat Lung
    Article Snippet: Paragraph title: Cloning and purification of recombinant NTHI TbpB. ... To purify rTbpB, cultures of BL21 [F− ompT hsd SB (rB − mB − ) gal dcm ] (Novagen, Madison, Wis.) containing pCU17 were grown in Luria-Bertani broth supplemented with 3.6 g of glucose per liter and 100 μg of ampicillin per ml.

    Article Title: Zinc Metalloproteinase ProA Directly Activates Legionella pneumophila PlaC Glycerophospholipid:cholesterol Acyltransferase *
    Article Snippet: .. E. coli DH5α or BL21 was used for the propagation of recombinant plasmid DNA with backbones of the following vectors: pMMB2002 , pet28a(+) (Novagen), pBADTOPO (Invitrogen), pBCKS (Stratagene), pGEX-6P-1 (GE Healthcare), pGEM-T Easy (Promega GmbH), and pET160 (Invitrogen). .. Genomic and plasmid DNA were prepared, amplified, and sequenced according to standard protocols.

    Article Title: Production of bioactive liver-targeting interferon Mu-IFN-CSP by soluble prokaryotic expression
    Article Snippet: .. E. coli strain DH5α (Novagen, Madison, USA) was employed for gene manipulation and BL21 (DE3; Novagen, Madison, USA) was employed to construct recombinant expression strain. .. Recombinant IFN-CSP/pET-21b expression plasmid was obtained from Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangzhou, People’s Republic of China.

    Mutagenesis:

    Article Title: Molecular basis for transfer RNA recognition by the double-stranded RNA-binding domain of human dihydrouridine synthase 2
    Article Snippet: Protein purification hDus2 or its dsRBD (T339-K451 construct) containing both the NTE and CTE was cloned in a pET11d vector between BamHI and NcoI and expressed in BL21(DE3)pLysS or BL21(DE3) respectively (Novagen) in LB medium and was purified as previously described ( ). .. Point mutations were carried out with Q5® Site-directed mutagenesis kit (New England BioLabs) following the recommended procedure and done on dsRDB-pET11d vector.

    Isolation:

    Article Title: Reduced Renal Colonization and Enhanced Protection by Leptospiral Factor H Binding Proteins as a Multisubunit Vaccine against Leptospirosis in Hamsters
    Article Snippet: Bacterial Strains and Culture Conditions Low-passage virulent Leptospira interrogans serovar Pomona (directly isolated from hamsters followed by < 5 in vitro passages) were used in all experiments [ ]. .. Escherichia coli strains DH5α and BL21 (DE3) pLysS (Novagen, Darmstadt, Germany) were grown at 37 °C in Luria–Bertani (LB) medium with the addition of 100 µg/mL ampicillin and 30 µg/mL chloramphenicol when required.

    Purification:

    Article Title: Immunization with Recombinant Transferrin Binding Protein B Enhances Clearance of Nontypeable Haemophilus influenzae from the Rat Lung
    Article Snippet: Paragraph title: Cloning and purification of recombinant NTHI TbpB. ... To purify rTbpB, cultures of BL21 [F− ompT hsd SB (rB − mB − ) gal dcm ] (Novagen, Madison, Wis.) containing pCU17 were grown in Luria-Bertani broth supplemented with 3.6 g of glucose per liter and 100 μg of ampicillin per ml.

    Article Title: The pCri System: A Vector Collection for Recombinant Protein Expression and Purification
    Article Snippet: DNA was purified from PCR reactions, enzymatic reactions, agarose gel band extractions, and vector extractions using OMEGA-Biotek purification kits. .. Chemically competent E. coli DH5α, BL21 (DE3), and Origami 2 (DE3) cells (Novagen) were prepared and transformed following Hanahan method .

    Article Title: Molecular basis for transfer RNA recognition by the double-stranded RNA-binding domain of human dihydrouridine synthase 2
    Article Snippet: .. Protein purification hDus2 or its dsRBD (T339-K451 construct) containing both the NTE and CTE was cloned in a pET11d vector between BamHI and NcoI and expressed in BL21(DE3)pLysS or BL21(DE3) respectively (Novagen) in LB medium and was purified as previously described ( ). .. Point mutations were carried out with Q5® Site-directed mutagenesis kit (New England BioLabs) following the recommended procedure and done on dsRDB-pET11d vector.

    Protein Purification:

    Article Title: Molecular basis for transfer RNA recognition by the double-stranded RNA-binding domain of human dihydrouridine synthase 2
    Article Snippet: .. Protein purification hDus2 or its dsRBD (T339-K451 construct) containing both the NTE and CTE was cloned in a pET11d vector between BamHI and NcoI and expressed in BL21(DE3)pLysS or BL21(DE3) respectively (Novagen) in LB medium and was purified as previously described ( ). .. Point mutations were carried out with Q5® Site-directed mutagenesis kit (New England BioLabs) following the recommended procedure and done on dsRDB-pET11d vector.

    Sequencing:

    Article Title: Zinc Metalloproteinase ProA Directly Activates Legionella pneumophila PlaC Glycerophospholipid:cholesterol Acyltransferase *
    Article Snippet: Paragraph title: DNA Techniques and Sequence Analysis ... E. coli DH5α or BL21 was used for the propagation of recombinant plasmid DNA with backbones of the following vectors: pMMB2002 , pet28a(+) (Novagen), pBADTOPO (Invitrogen), pBCKS (Stratagene), pGEX-6P-1 (GE Healthcare), pGEM-T Easy (Promega GmbH), and pET160 (Invitrogen).

    Lysis:

    Article Title: Preparation of Biologically Active Single-Chain Variable Antibody Fragments that Target the HIV-1 gp120 V3 Loop
    Article Snippet: To optimize scFv expression in various E. coli expression strains, including BL21 (DE3), Rosetta 2 (DE3) (Novagen), BL21 Gold (DE3) pLysS (Stratagene), and BL21 Star (DE3) (Invitrogen), a similar protocol is used with modifications of the growing culture (in LB with the appropriate antibiotics) at 37 °C and inducing expression with 0.5 mM IPTG. .. Each frozen cell pellet was thawed on ice and resuspended in 5 ml lysis buffer containing 50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1 mM EDTA, 0.1% Triton X-100, 1 mM phenylmethylsulfonyl fluoride (PMSF), and 250 μg/ml lysozyme.

    Plasmid Preparation:

    Article Title: Expression and purification of toxic anti-breast cancer p28-NRC chimeric protein
    Article Snippet: Bacterial strain, plasmids, and culture conditions E. coli strains TOP10, BL21(DE3) and BL21(DE3)pLysS, and pET-28a and pET-32a plasmids were obtained from Novagen (Madison, WI). .. KpnI , XhoI , and NdeI restriction enzymes, T4 DNA Ligase, GeneJet Gel Extraction kit, and GenJet plasmid Preparation kit were obtained from Thermo scientific (MD, USA).

    Article Title: Immunization with Recombinant Transferrin Binding Protein B Enhances Clearance of Nontypeable Haemophilus influenzae from the Rat Lung
    Article Snippet: This plasmid is engineered to express recombinant TbpB as a glutathione S -transferase (GST) fusion protein with a thrombin cleavage recognition site between the two proteins. .. To purify rTbpB, cultures of BL21 [F− ompT hsd SB (rB − mB − ) gal dcm ] (Novagen, Madison, Wis.) containing pCU17 were grown in Luria-Bertani broth supplemented with 3.6 g of glucose per liter and 100 μg of ampicillin per ml.

    Article Title: Zinc Metalloproteinase ProA Directly Activates Legionella pneumophila PlaC Glycerophospholipid:cholesterol Acyltransferase *
    Article Snippet: .. E. coli DH5α or BL21 was used for the propagation of recombinant plasmid DNA with backbones of the following vectors: pMMB2002 , pet28a(+) (Novagen), pBADTOPO (Invitrogen), pBCKS (Stratagene), pGEX-6P-1 (GE Healthcare), pGEM-T Easy (Promega GmbH), and pET160 (Invitrogen). .. Genomic and plasmid DNA were prepared, amplified, and sequenced according to standard protocols.

    Article Title: Production of bioactive liver-targeting interferon Mu-IFN-CSP by soluble prokaryotic expression
    Article Snippet: Plasmids and strains pMD20-T (Takara, Otsu, Japan) was employed to clone gene and pET-21b (Novagen, Madison, USA) was employed to construct expression plasmid. .. E. coli strain DH5α (Novagen, Madison, USA) was employed for gene manipulation and BL21 (DE3; Novagen, Madison, USA) was employed to construct recombinant expression strain.

    Article Title: Soluble Expression of Humanized Anti-CD20 Single Chain Antibody in Escherichia coli by Cytoplasmic Chaperones Co-expression
    Article Snippet: .. Bacterial strains, media, and plasmids E. coli DH5α, BL21 (DE3) and expression vector pET22b (+) were purchased from Novagen. .. IPTG, Taq DNA polymerase was obtained from Promega Company.

    Article Title: YeeV is an Escherichia coli Toxin that Inhibits Cell Division by Targeting the Cytoskeleton Proteins, FtsZ and MreB
    Article Snippet: E. coli BW25113 ( ΔaraBD ) (Datsenko and Wanner, 2000), BL21 (DE3) were grown in M9 medium at 37° C. Plasmids pET-28a- yeeV , pET-28a- ftsZ , pET-28a- mreB and pET-28a -sulA were constructed in pET-28a (Novagen) with an N-terminal His6 tag. .. To generate the N-terminal GFP-fusion plasmids, a DNA fragment of gfp from plasmid pAcGFP1 (Clontech laboratories, Inc.) was cloned into pBAD33 to generate pBAD33- gfp.

    Article Title: The pCri System: A Vector Collection for Recombinant Protein Expression and Purification
    Article Snippet: Paragraph title: Genetic manipulations and vector preparation ... Chemically competent E. coli DH5α, BL21 (DE3), and Origami 2 (DE3) cells (Novagen) were prepared and transformed following Hanahan method .

    Article Title: Soluble Expression of Humanized Anti-CD20 Single Chain Antibody in Escherichia coli by Cytoplasmic Chaperones Co-expression
    Article Snippet: .. E. coli DH5α, BL21 (DE3) and expression vector pET22b (+) were purchased from Novagen. .. IPTG, Taq DNA polymerase was obtained from Promega Company.

    Article Title: Host cell and expression engineering for development of an E. coli ketoreductase catalyst: Enhancement of formate dehydrogenase activity for regeneration of NADH
    Article Snippet: Strains and plasmids The E. coli strains used were JM109 from Promega (Madison, WI, USA), BL21 (DE3) and Rosetta 2 (DE3) from Novagen (VWR International GmbH, Vienna, Austria) and BL21 star (DE3) from Invitrogen (Carlsbad, California, USA). .. The construction of the E. coli BL21 (DE3) (BL21_XR_FDH) harbouring Ct XR and Cb FDH genes on a pETDuet-1 vector (pETDuet_XR_FDH) was described elsewhere [ ].

    Article Title: Molecular basis for transfer RNA recognition by the double-stranded RNA-binding domain of human dihydrouridine synthase 2
    Article Snippet: .. Protein purification hDus2 or its dsRBD (T339-K451 construct) containing both the NTE and CTE was cloned in a pET11d vector between BamHI and NcoI and expressed in BL21(DE3)pLysS or BL21(DE3) respectively (Novagen) in LB medium and was purified as previously described ( ). .. Point mutations were carried out with Q5® Site-directed mutagenesis kit (New England BioLabs) following the recommended procedure and done on dsRDB-pET11d vector.

    Positron Emission Tomography:

    Article Title: Expression and purification of toxic anti-breast cancer p28-NRC chimeric protein
    Article Snippet: .. Bacterial strain, plasmids, and culture conditions E. coli strains TOP10, BL21(DE3) and BL21(DE3)pLysS, and pET-28a and pET-32a plasmids were obtained from Novagen (Madison, WI). .. KpnI , XhoI , and NdeI restriction enzymes, T4 DNA Ligase, GeneJet Gel Extraction kit, and GenJet plasmid Preparation kit were obtained from Thermo scientific (MD, USA).

    Article Title: Production of bioactive liver-targeting interferon Mu-IFN-CSP by soluble prokaryotic expression
    Article Snippet: Plasmids and strains pMD20-T (Takara, Otsu, Japan) was employed to clone gene and pET-21b (Novagen, Madison, USA) was employed to construct expression plasmid. .. E. coli strain DH5α (Novagen, Madison, USA) was employed for gene manipulation and BL21 (DE3; Novagen, Madison, USA) was employed to construct recombinant expression strain.

    Article Title: YeeV is an Escherichia coli Toxin that Inhibits Cell Division by Targeting the Cytoskeleton Proteins, FtsZ and MreB
    Article Snippet: .. E. coli BW25113 ( ΔaraBD ) (Datsenko and Wanner, 2000), BL21 (DE3) were grown in M9 medium at 37° C. Plasmids pET-28a- yeeV , pET-28a- ftsZ , pET-28a- mreB and pET-28a -sulA were constructed in pET-28a (Novagen) with an N-terminal His6 tag. .. Plasmids pBAD33- yeeV and pBAD33- sulA were constructed from pBAD33 (Guzman et al., 1995) to tightly regulate gene expression by the addition of arabinose.

    Article Title: Construction of a Novel Liver-Targeting Fusion Interferon by Incorporation of a Plasmodium Region I-Plus Peptide
    Article Snippet: .. Bacterial Strains, Plasmids, and Culture Media E. coli strain DH5α (Novagen, USA) and BL21 (DE3; Novagen, USA) were used as the host for gene manipulation and expression of fusion protein, respectively. pMD20-T (Takara, Japan) and pET-21b (Novagen, USA) were applied for gene cloning and expression, respectively. ..

    Agarose Gel Electrophoresis:

    Article Title: Immunization with Recombinant Transferrin Binding Protein B Enhances Clearance of Nontypeable Haemophilus influenzae from the Rat Lung
    Article Snippet: Agarose gel analysis of the PCR product demonstrated a ∼1.8 kb fragment, which was purified with Bresaclean resin (Bresatec Ltd. Adelaide, Australia), digested with Bam HI, and cloned into the Bam HI restriction sites in plasmid pGEX2T (Pharmacia Biotech, Uppsala, Sweden) to produce plasmid pCU17. .. To purify rTbpB, cultures of BL21 [F− ompT hsd SB (rB − mB − ) gal dcm ] (Novagen, Madison, Wis.) containing pCU17 were grown in Luria-Bertani broth supplemented with 3.6 g of glucose per liter and 100 μg of ampicillin per ml.

    Article Title: The pCri System: A Vector Collection for Recombinant Protein Expression and Purification
    Article Snippet: DNA was purified from PCR reactions, enzymatic reactions, agarose gel band extractions, and vector extractions using OMEGA-Biotek purification kits. .. Chemically competent E. coli DH5α, BL21 (DE3), and Origami 2 (DE3) cells (Novagen) were prepared and transformed following Hanahan method .

    In Vitro:

    Article Title: Reduced Renal Colonization and Enhanced Protection by Leptospiral Factor H Binding Proteins as a Multisubunit Vaccine against Leptospirosis in Hamsters
    Article Snippet: Bacterial Strains and Culture Conditions Low-passage virulent Leptospira interrogans serovar Pomona (directly isolated from hamsters followed by < 5 in vitro passages) were used in all experiments [ ]. .. Escherichia coli strains DH5α and BL21 (DE3) pLysS (Novagen, Darmstadt, Germany) were grown at 37 °C in Luria–Bertani (LB) medium with the addition of 100 µg/mL ampicillin and 30 µg/mL chloramphenicol when required.

    Incubation:

    Article Title: Preparation of Biologically Active Single-Chain Variable Antibody Fragments that Target the HIV-1 gp120 V3 Loop
    Article Snippet: The remaining culture was incubated at 37 °C with shaking for three hours without addition of Isopropyl β-D-1-thiogalactopyranoside (IPTG). .. To optimize scFv expression in various E. coli expression strains, including BL21 (DE3), Rosetta 2 (DE3) (Novagen), BL21 Gold (DE3) pLysS (Stratagene), and BL21 Star (DE3) (Invitrogen), a similar protocol is used with modifications of the growing culture (in LB with the appropriate antibiotics) at 37 °C and inducing expression with 0.5 mM IPTG.

    Concentration Assay:

    Article Title: Preparation of Biologically Active Single-Chain Variable Antibody Fragments that Target the HIV-1 gp120 V3 Loop
    Article Snippet: Cultures in the three other flasks were induced with IPTG at final concentration of 0.25 mM, 0.5 mM, and 1 mM respectively and continue shaking at 37 °C for three hours. .. To optimize scFv expression in various E. coli expression strains, including BL21 (DE3), Rosetta 2 (DE3) (Novagen), BL21 Gold (DE3) pLysS (Stratagene), and BL21 Star (DE3) (Invitrogen), a similar protocol is used with modifications of the growing culture (in LB with the appropriate antibiotics) at 37 °C and inducing expression with 0.5 mM IPTG.

    DNA Purification:

    Article Title: The pCri System: A Vector Collection for Recombinant Protein Expression and Purification
    Article Snippet: When necessary, a second round of digestion was performed before the final DNA purification step. .. Chemically competent E. coli DH5α, BL21 (DE3), and Origami 2 (DE3) cells (Novagen) were prepared and transformed following Hanahan method .

    Gel Extraction:

    Article Title: Expression and purification of toxic anti-breast cancer p28-NRC chimeric protein
    Article Snippet: Bacterial strain, plasmids, and culture conditions E. coli strains TOP10, BL21(DE3) and BL21(DE3)pLysS, and pET-28a and pET-32a plasmids were obtained from Novagen (Madison, WI). .. KpnI , XhoI , and NdeI restriction enzymes, T4 DNA Ligase, GeneJet Gel Extraction kit, and GenJet plasmid Preparation kit were obtained from Thermo scientific (MD, USA).

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