e coli bl21  (Millipore)


Bioz Verified Symbol Millipore is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99

    Structured Review

    Millipore e coli bl21
    E Coli Bl21, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 792 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli bl21/product/Millipore
    Average 99 stars, based on 792 article reviews
    Price from $9.99 to $1999.99
    e coli bl21 - by Bioz Stars, 2020-01
    99/100 stars

    Images

    Related Articles

    Clone Assay:

    Article Title: Functional Characterization by Genetic Complementation of aroB-Encoded Dehydroquinate Synthase from Mycobacterium tuberculosis H37Rv and Its Heterologous Expression and Purification ▿
    Article Snippet: Paragraph title: Amplification, cloning, and overexpression of the M. tuberculosis aroB gene. ... The recombinant pET23a(+):: aroB plasmid was introduced into Escherichia coli BL21(DE3) (Novagen) electrocompetent cells and selected on LB agar plates containing 50 μg ml−1 carbenicillin.

    Article Title: Cofactor-induced reversible folding of Flavodoxin-4 from Lactobacillus acidophilus
    Article Snippet: The cloning junctions were confirmed by DNA sequencing. .. Expression was performed in E. Coli BL21(DE3) cells (EMD Biosciences).

    Article Title: Molecular characterization and expression analysis of iron superoxide dismutase gene from Pseudochlorella pringsheimii (Trebouxiophyceae, Chlorophyta)
    Article Snippet: The previous product (644 bp, Fig. a) was gel-purified, and cloned into the pET28c expression vector (Novagen, USA). .. The FeSOD nucleotide sequences in the expression vector were sequenced again for validation, and then the vector was transformed into E. coli BL21(DE3)pLysS cells (Novagen, USA).

    Article Title: A Novel Prokaryote-Type ECF/ABC Transporter Module in Chloroplast Metal Homeostasis
    Article Snippet: Antiserum Production To raise antisera against At-ABCI10 and At-ABCI11, cDNA was PCR-amplified on the respective SALK pUNI clones U62184 and U51365 ( ). .. The resulting mature versions of At-ABCI10 and At-ABCI11 were subcloned into the pET21d (Novagen) plasmid vector and used for overexpression after transforming E. coli BL21(DE3) cells (Novagen).

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: The cloned nucleotide sequence was confirmed by sequencing and subcloned into the NdeI site in the multiple cloning site of the pET-16b vector (Novagen) to fuse a His10 tag sequence to the N terminus of the neutral SMase 1 open reading frame. .. Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1.

    Article Title: A post-translational modification signature defines changes in soluble tau correlating with oligomerization in early stage Alzheimer’s disease brain
    Article Snippet: Recombinant tau protein purification Tau variants (full length protein and a fragment encoding amino acids 256–368) were cloned into the pET19b vector (Novagen) in between the NcoI and BamHI restriction sites. .. The pET19b-Tau plasmids were transformed into E. coli BL21(DE3) cells (Novagen).

    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: Cofactor-induced reversible folding of Flavodoxin-4 from Lactobacillus acidophilus
    Article Snippet: Expression was performed in E. Coli BL21(DE3) cells (EMD Biosciences). .. The temperature was then reduced to 18°C, and protein expression was induced by 1 m M isopropyl β - d −1-thiogalactopyranoside for 20 h. Cells were harvested by centrifugation at 5,000 g at 4°C for 5 min and re-suspended in buffer A (20 m M sodium phosphate at pH 7.4, 200 m M NaCl, 20 m M imidazole) supplemented with Complete EDTA-free protease inhibitor cocktail tablets (Roche).

    Article Title: Effects of allelic variations in the human myxovirus resistance protein A on its antiviral activity
    Article Snippet: Human MxA and the indicated mutants/variants were expressed as N-terminal His6 -tagged fusions followed by a PreScissionTM cleavage site from a pET28 plasmid in the E. coli BL21 ( DE3 ) Rosetta strain (Novagen) as described by Gao et al . ( ). .. Following centrifugation, bacterial pellets were resuspended in ice-cold 50 m m HEPES (pH 7.5), 800 m m NaCl, 30 m m imidazole, 5 m m MgCl2 , 1 μ m DNase I, 2.5 m m β-mercaptoethanol (β-ME), 500 μ m Pefabloc SC (Roth) and lysed in a microfluidizer (Microfluidics).

    Article Title: A Novel Prokaryote-Type ECF/ABC Transporter Module in Chloroplast Metal Homeostasis
    Article Snippet: The resulting mature versions of At-ABCI10 and At-ABCI11 were subcloned into the pET21d (Novagen) plasmid vector and used for overexpression after transforming E. coli BL21(DE3) cells (Novagen). .. Afterwards, pelleted cells (4°C, 6,000g, 15 min) were resuspended in cell lysis solution (50 mM Tris-HCl, pH 8.0, 25% [w/v] sucrose, 1 mM EDTA, 100 mg/ml DNase) and sonicated three times for 30 s. Inclusion bodies were collected by centrifugation at 4°C and 20,000g for 30 min.

    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).

    Article Title: Generation of a recombinant Aggregatibacter actinomycetemcomitans RTX toxin in Escherichia coli
    Article Snippet: Plasmids were transformed into E. coli BL21(λDE3) cells (Novagen, Madison, WI). .. The cell pellet was collected by centrifugation at 15,000 × g for 10 min, suspended in 8 ml 20 mM Tris, 250 mM NaCl and 200 μM CaCl2 pH 6.8 buffer containing protease inhibitors cocktail (Sigma-Aldrich, St. Louis, MO) and the cells were disrupted by sonication on ice for 6 cycles of 30 min on/30 min off using Virtis Virsonic 475 Sonicator (SP Scientific, Gardiner, NY).

    Amplification:

    Article Title: Functional Characterization by Genetic Complementation of aroB-Encoded Dehydroquinate Synthase from Mycobacterium tuberculosis H37Rv and Its Heterologous Expression and Purification ▿
    Article Snippet: Paragraph title: Amplification, cloning, and overexpression of the M. tuberculosis aroB gene. ... The recombinant pET23a(+):: aroB plasmid was introduced into Escherichia coli BL21(DE3) (Novagen) electrocompetent cells and selected on LB agar plates containing 50 μg ml−1 carbenicillin.

    Article Title: Cofactor-induced reversible folding of Flavodoxin-4 from Lactobacillus acidophilus
    Article Snippet: V-PIPE and I-PIPE PCR products were mixed, and the amplified DNA fragments were annealed. .. Expression was performed in E. Coli BL21(DE3) cells (EMD Biosciences).

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: Preparation of Recombinant Neutral SMase —The neutral SMase 1 cDNA was amplified by PCR using sense primer 5′-CATATGGCACCACAGCAGCCCGGCAAACTG-3′ and antisense primer 5′-CATATGTTATTCCTCCCGTTTGAAAGGACT-3′, each containing an NdeI site; the amplimer was subcloned into the pGEM-T Easy plasmid vector by the TA cloning method (Promega). .. Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1.

    Synthesized:

    Article Title: Functional Characterization by Genetic Complementation of aroB-Encoded Dehydroquinate Synthase from Mycobacterium tuberculosis H37Rv and Its Heterologous Expression and Purification ▿
    Article Snippet: Two oligonucleotides (5′-GGC CATATG ACCGATATCGGCGCACCCG-3′ and 5′-A GGATCC TCATGGGGCGCAAACTCCGGC-3′) complementary to the amino-terminal coding and carboxy-terminal noncoding strands of the M. tuberculosis aroB gene ( ) were synthesized to contain, respectively, NdeI and BamHI restriction sites (underlined). .. The recombinant pET23a(+):: aroB plasmid was introduced into Escherichia coli BL21(DE3) (Novagen) electrocompetent cells and selected on LB agar plates containing 50 μg ml−1 carbenicillin.

    Article Title: Dystrophin As A Molecular Shock Absorber
    Article Snippet: Four fragments of the dystrophin central rod domain ( i.e ., SR01–05: 338th -938th ; SR06–10: 939th -1466th ; SR11–17: 1464th -2210th ; SR18–24: 2209th -3044th ) were synthesized by PCR using plasmid template containing the full-length human dystrophin sequence. .. Each of the resulting plasmids was co-transformed with a BirA plasmid and expressed in Escherichia coli BL21 (DE3) cultured in LB-media with D-Biotin (Sigma Aldrich), and affinity purified through His-tag.

    TA Cloning:

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: Preparation of Recombinant Neutral SMase —The neutral SMase 1 cDNA was amplified by PCR using sense primer 5′-CATATGGCACCACAGCAGCCCGGCAAACTG-3′ and antisense primer 5′-CATATGTTATTCCTCCCGTTTGAAAGGACT-3′, each containing an NdeI site; the amplimer was subcloned into the pGEM-T Easy plasmid vector by the TA cloning method (Promega). .. Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1.

    Construct:

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: This construct was named pETZNSMase 1. .. Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1.

    Article Title: Dystrophin As A Molecular Shock Absorber
    Article Snippet: Paragraph title: Plasmids constructs and protein expression. ... Each of the resulting plasmids was co-transformed with a BirA plasmid and expressed in Escherichia coli BL21 (DE3) cultured in LB-media with D-Biotin (Sigma Aldrich), and affinity purified through His-tag.

    Electrophoresis:

    Article Title: Functional Characterization by Genetic Complementation of aroB-Encoded Dehydroquinate Synthase from Mycobacterium tuberculosis H37Rv and Its Heterologous Expression and Purification ▿
    Article Snippet: The PCR product (1,089 bp) was purified by electrophoresis, digested with NdeI and BamHI (Boehringer Mannheim), and ligated into a pET23a(+) expression vector (Novagen) that had previously been digested with the same restriction enzymes. .. The recombinant pET23a(+):: aroB plasmid was introduced into Escherichia coli BL21(DE3) (Novagen) electrocompetent cells and selected on LB agar plates containing 50 μg ml−1 carbenicillin.

    Incubation:

    Article Title: Molecular characterization and expression analysis of iron superoxide dismutase gene from Pseudochlorella pringsheimii (Trebouxiophyceae, Chlorophyta)
    Article Snippet: The FeSOD nucleotide sequences in the expression vector were sequenced again for validation, and then the vector was transformed into E. coli BL21(DE3)pLysS cells (Novagen, USA). .. After the E. coli cells reached a cell-density of OD600 = 0.6, the expression of the FeSOD protein was induced by the addition of 25 μM IPTG (isopropyl-β-D-thiogalactopyranoside), followed by incubation at 28 °C for 15 h. Purification of the expressed protein was done via affinity chromatography on a HiTrap HP column (GE Healthcare, USA), quantified by A280 and Bradford ( ) method (Biorad, USA) and the purity was validated by 12% SDS-PAGE (SDS–polyacrylamide gel electrophoresis; Laemmli , Fig. b).

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1. .. The culture was transferred to 1000 ml of fresh LB medium with 100 μg/ml ampicillin in a 5-liter flask, and the above incubation conditions were continued until turbidity at 600 nm reached 0.8.

    GPC Assay:

    Article Title: The RNase YbeY Is Vital for Ribosome Maturation, Stress Resistance, and Virulence of the Natural Genetic Engineer Agrobacterium tumefaciens
    Article Snippet: Hfq6×His and YbeY6×His fusion proteins were expressed in E. coli BL21(DE3) cells, using pET-plasmid-based expression from a T7 promoter (Novagen, Madison, WI). .. To assess the oligomeric state, we performed gel permeation chromatography.

    Expressing:

    Article Title: Complete reconstitution of the diverse pathways of gentamicin B biosynthesis
    Article Snippet: .. For expression of the recombinant proteins (GenM1, GenD2, GenS2, GenN, GenD1, GenQ, GenB1, GenJ, and GenK2), the E. coli BL21(DE3) (Novagen), E. coli BL21(DE3)pLysS (Novagen), E. coli Rosetta gamiTM2 (DE3) (Novagen) and E. coli ArcticExpress(DE3) (Agilent technologies, Santa Clara, CA, USA) were used as heterologous hosts. ..

    Article Title: Functional Characterization by Genetic Complementation of aroB-Encoded Dehydroquinate Synthase from Mycobacterium tuberculosis H37Rv and Its Heterologous Expression and Purification ▿
    Article Snippet: The PCR product (1,089 bp) was purified by electrophoresis, digested with NdeI and BamHI (Boehringer Mannheim), and ligated into a pET23a(+) expression vector (Novagen) that had previously been digested with the same restriction enzymes. .. The recombinant pET23a(+):: aroB plasmid was introduced into Escherichia coli BL21(DE3) (Novagen) electrocompetent cells and selected on LB agar plates containing 50 μg ml−1 carbenicillin.

    Article Title: Cofactor-induced reversible folding of Flavodoxin-4 from Lactobacillus acidophilus
    Article Snippet: .. Expression was performed in E. Coli BL21(DE3) cells (EMD Biosciences). ..

    Article Title: Molecular characterization and expression analysis of iron superoxide dismutase gene from Pseudochlorella pringsheimii (Trebouxiophyceae, Chlorophyta)
    Article Snippet: .. The FeSOD nucleotide sequences in the expression vector were sequenced again for validation, and then the vector was transformed into E. coli BL21(DE3)pLysS cells (Novagen, USA). .. After the E. coli cells reached a cell-density of OD600 = 0.6, the expression of the FeSOD protein was induced by the addition of 25 μM IPTG (isopropyl-β-D-thiogalactopyranoside), followed by incubation at 28 °C for 15 h. Purification of the expressed protein was done via affinity chromatography on a HiTrap HP column (GE Healthcare, USA), quantified by A280 and Bradford ( ) method (Biorad, USA) and the purity was validated by 12% SDS-PAGE (SDS–polyacrylamide gel electrophoresis; Laemmli , Fig. b).

    Article Title: Effects of allelic variations in the human myxovirus resistance protein A on its antiviral activity
    Article Snippet: Paragraph title: Protein expression and purification ... Human MxA and the indicated mutants/variants were expressed as N-terminal His6 -tagged fusions followed by a PreScissionTM cleavage site from a pET28 plasmid in the E. coli BL21 ( DE3 ) Rosetta strain (Novagen) as described by Gao et al . ( ).

    Article Title: The RNase YbeY Is Vital for Ribosome Maturation, Stress Resistance, and Virulence of the Natural Genetic Engineer Agrobacterium tumefaciens
    Article Snippet: .. Hfq6×His and YbeY6×His fusion proteins were expressed in E. coli BL21(DE3) cells, using pET-plasmid-based expression from a T7 promoter (Novagen, Madison, WI). .. To assess the oligomeric state, we performed gel permeation chromatography.

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1. .. Isopropyl-1-thio-β- d -galactopyranoside was added to a final concentration of 1 m m , and the culture was grown for an additional 4 h to induce the expression of the transgene.

    Article Title: A post-translational modification signature defines changes in soluble tau correlating with oligomerization in early stage Alzheimer’s disease brain
    Article Snippet: The pET19b-Tau plasmids were transformed into E. coli BL21(DE3) cells (Novagen). .. The expression of the tau proteins was induced by the addition of 1 mM IPTG.

    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: Generation of a recombinant Aggregatibacter actinomycetemcomitans RTX toxin in Escherichia coli
    Article Snippet: Paragraph title: His-tagged LtxA expression and purification ... Plasmids were transformed into E. coli BL21(λDE3) cells (Novagen, Madison, WI).

    Article Title: Dystrophin As A Molecular Shock Absorber
    Article Snippet: Paragraph title: Plasmids constructs and protein expression. ... Each of the resulting plasmids was co-transformed with a BirA plasmid and expressed in Escherichia coli BL21 (DE3) cultured in LB-media with D-Biotin (Sigma Aldrich), and affinity purified through His-tag.

    Transformation Assay:

    Article Title: Cofactor-induced reversible folding of Flavodoxin-4 from Lactobacillus acidophilus
    Article Snippet: E. coli GeneHogs (Invitrogen) competent cells were transformed with the I-PIPE/V-PIPE mixture and dispensed on selective LB-agar plates. .. Expression was performed in E. Coli BL21(DE3) cells (EMD Biosciences).

    Article Title: Molecular characterization and expression analysis of iron superoxide dismutase gene from Pseudochlorella pringsheimii (Trebouxiophyceae, Chlorophyta)
    Article Snippet: .. The FeSOD nucleotide sequences in the expression vector were sequenced again for validation, and then the vector was transformed into E. coli BL21(DE3)pLysS cells (Novagen, USA). .. After the E. coli cells reached a cell-density of OD600 = 0.6, the expression of the FeSOD protein was induced by the addition of 25 μM IPTG (isopropyl-β-D-thiogalactopyranoside), followed by incubation at 28 °C for 15 h. Purification of the expressed protein was done via affinity chromatography on a HiTrap HP column (GE Healthcare, USA), quantified by A280 and Bradford ( ) method (Biorad, USA) and the purity was validated by 12% SDS-PAGE (SDS–polyacrylamide gel electrophoresis; Laemmli , Fig. b).

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: .. Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1. ..

    Article Title: Transferrin Receptor 1-Associated Iron Accumulation and Oxidative Stress Provides a Way for Grass Carp to Fight against Reovirus Infection
    Article Snippet: .. To address this, pGEX-4T-1-TfR1 and pET32a(+)-Tf plasmids were transformed into the Escherichia coli BL21 and BL21(DE3) cells (Novagen), respectively. .. Then, CiTfR1 and CiTf proteins were expressed in vitro and purified.

    Article Title: A post-translational modification signature defines changes in soluble tau correlating with oligomerization in early stage Alzheimer’s disease brain
    Article Snippet: .. The pET19b-Tau plasmids were transformed into E. coli BL21(DE3) cells (Novagen). ..

    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: Generation of a recombinant Aggregatibacter actinomycetemcomitans RTX toxin in Escherichia coli
    Article Snippet: .. Plasmids were transformed into E. coli BL21(λDE3) cells (Novagen, Madison, WI). ..

    Over Expression:

    Article Title: Functional Characterization by Genetic Complementation of aroB-Encoded Dehydroquinate Synthase from Mycobacterium tuberculosis H37Rv and Its Heterologous Expression and Purification ▿
    Article Snippet: Paragraph title: Amplification, cloning, and overexpression of the M. tuberculosis aroB gene. ... The recombinant pET23a(+):: aroB plasmid was introduced into Escherichia coli BL21(DE3) (Novagen) electrocompetent cells and selected on LB agar plates containing 50 μg ml−1 carbenicillin.

    Article Title: A Novel Prokaryote-Type ECF/ABC Transporter Module in Chloroplast Metal Homeostasis
    Article Snippet: .. The resulting mature versions of At-ABCI10 and At-ABCI11 were subcloned into the pET21d (Novagen) plasmid vector and used for overexpression after transforming E. coli BL21(DE3) cells (Novagen). ..

    Transfection:

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: At 24 h after transfection, the cells were washed twice with PBS and homogenized in lysis buffer for neutral SMase assay. .. Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1.

    Chromatography:

    Article Title: The RNase YbeY Is Vital for Ribosome Maturation, Stress Resistance, and Virulence of the Natural Genetic Engineer Agrobacterium tumefaciens
    Article Snippet: Hfq6×His and YbeY6×His fusion proteins were expressed in E. coli BL21(DE3) cells, using pET-plasmid-based expression from a T7 promoter (Novagen, Madison, WI). .. After Ni-NTA chromatography, purified His-tagged fusion proteins were separated by size via a Superdex 200 10/300 GL column (HEPES buffer [pH 7.5]) on an ÄKTA system (GE Healthcare).

    Protease Inhibitor:

    Article Title: Cofactor-induced reversible folding of Flavodoxin-4 from Lactobacillus acidophilus
    Article Snippet: Expression was performed in E. Coli BL21(DE3) cells (EMD Biosciences). .. The temperature was then reduced to 18°C, and protein expression was induced by 1 m M isopropyl β - d −1-thiogalactopyranoside for 20 h. Cells were harvested by centrifugation at 5,000 g at 4°C for 5 min and re-suspended in buffer A (20 m M sodium phosphate at pH 7.4, 200 m M NaCl, 20 m M imidazole) supplemented with Complete EDTA-free protease inhibitor cocktail tablets (Roche).

    Cell Culture:

    Article Title: Dystrophin As A Molecular Shock Absorber
    Article Snippet: .. Each of the resulting plasmids was co-transformed with a BirA plasmid and expressed in Escherichia coli BL21 (DE3) cultured in LB-media with D-Biotin (Sigma Aldrich), and affinity purified through His-tag. .. A vertical magnetic tweezers setup, , was combined with a disturbance-free, rapid solution-exchange flow channel for conducting in vitro protein stretching experiments.

    DNA Sequencing:

    Article Title: Cofactor-induced reversible folding of Flavodoxin-4 from Lactobacillus acidophilus
    Article Snippet: The cloning junctions were confirmed by DNA sequencing. .. Expression was performed in E. Coli BL21(DE3) cells (EMD Biosciences).

    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.

    Polymerase Chain Reaction:

    Article Title: Functional Characterization by Genetic Complementation of aroB-Encoded Dehydroquinate Synthase from Mycobacterium tuberculosis H37Rv and Its Heterologous Expression and Purification ▿
    Article Snippet: The DNA sequence of the M. tuberculosis aroB gene was determined in order to confirm the identity, integrity, and absence of PCR-introduced mutations in the cloned gene. .. The recombinant pET23a(+):: aroB plasmid was introduced into Escherichia coli BL21(DE3) (Novagen) electrocompetent cells and selected on LB agar plates containing 50 μg ml−1 carbenicillin.

    Article Title: Cofactor-induced reversible folding of Flavodoxin-4 from Lactobacillus acidophilus
    Article Snippet: V-PIPE and I-PIPE PCR products were mixed, and the amplified DNA fragments were annealed. .. Expression was performed in E. Coli BL21(DE3) cells (EMD Biosciences).

    Article Title: A Novel Prokaryote-Type ECF/ABC Transporter Module in Chloroplast Metal Homeostasis
    Article Snippet: Antiserum Production To raise antisera against At-ABCI10 and At-ABCI11, cDNA was PCR-amplified on the respective SALK pUNI clones U62184 and U51365 ( ). .. The resulting mature versions of At-ABCI10 and At-ABCI11 were subcloned into the pET21d (Novagen) plasmid vector and used for overexpression after transforming E. coli BL21(DE3) cells (Novagen).

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: Preparation of Recombinant Neutral SMase —The neutral SMase 1 cDNA was amplified by PCR using sense primer 5′-CATATGGCACCACAGCAGCCCGGCAAACTG-3′ and antisense primer 5′-CATATGTTATTCCTCCCGTTTGAAAGGACT-3′, each containing an NdeI site; the amplimer was subcloned into the pGEM-T Easy plasmid vector by the TA cloning method (Promega). .. Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1.

    Article Title: Dystrophin As A Molecular Shock Absorber
    Article Snippet: Four fragments of the dystrophin central rod domain ( i.e ., SR01–05: 338th -938th ; SR06–10: 939th -1466th ; SR11–17: 1464th -2210th ; SR18–24: 2209th -3044th ) were synthesized by PCR using plasmid template containing the full-length human dystrophin sequence. .. Each of the resulting plasmids was co-transformed with a BirA plasmid and expressed in Escherichia coli BL21 (DE3) cultured in LB-media with D-Biotin (Sigma Aldrich), and affinity purified through His-tag.

    Sonication:

    Article Title: A Novel Prokaryote-Type ECF/ABC Transporter Module in Chloroplast Metal Homeostasis
    Article Snippet: The resulting mature versions of At-ABCI10 and At-ABCI11 were subcloned into the pET21d (Novagen) plasmid vector and used for overexpression after transforming E. coli BL21(DE3) cells (Novagen). .. Afterwards, pelleted cells (4°C, 6,000g, 15 min) were resuspended in cell lysis solution (50 mM Tris-HCl, pH 8.0, 25% [w/v] sucrose, 1 mM EDTA, 100 mg/ml DNase) and sonicated three times for 30 s. Inclusion bodies were collected by centrifugation at 4°C and 20,000g for 30 min.

    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).

    Article Title: Generation of a recombinant Aggregatibacter actinomycetemcomitans RTX toxin in Escherichia coli
    Article Snippet: Plasmids were transformed into E. coli BL21(λDE3) cells (Novagen, Madison, WI). .. The cell pellet was collected by centrifugation at 15,000 × g for 10 min, suspended in 8 ml 20 mM Tris, 250 mM NaCl and 200 μM CaCl2 pH 6.8 buffer containing protease inhibitors cocktail (Sigma-Aldrich, St. Louis, MO) and the cells were disrupted by sonication on ice for 6 cycles of 30 min on/30 min off using Virtis Virsonic 475 Sonicator (SP Scientific, Gardiner, NY).

    Affinity Purification:

    Article Title: Dystrophin As A Molecular Shock Absorber
    Article Snippet: .. Each of the resulting plasmids was co-transformed with a BirA plasmid and expressed in Escherichia coli BL21 (DE3) cultured in LB-media with D-Biotin (Sigma Aldrich), and affinity purified through His-tag. .. A vertical magnetic tweezers setup, , was combined with a disturbance-free, rapid solution-exchange flow channel for conducting in vitro protein stretching experiments.

    Recombinant:

    Article Title: Complete reconstitution of the diverse pathways of gentamicin B biosynthesis
    Article Snippet: .. For expression of the recombinant proteins (GenM1, GenD2, GenS2, GenN, GenD1, GenQ, GenB1, GenJ, and GenK2), the E. coli BL21(DE3) (Novagen), E. coli BL21(DE3)pLysS (Novagen), E. coli Rosetta gamiTM2 (DE3) (Novagen) and E. coli ArcticExpress(DE3) (Agilent technologies, Santa Clara, CA, USA) were used as heterologous hosts. ..

    Article Title: Functional Characterization by Genetic Complementation of aroB-Encoded Dehydroquinate Synthase from Mycobacterium tuberculosis H37Rv and Its Heterologous Expression and Purification ▿
    Article Snippet: .. The recombinant pET23a(+):: aroB plasmid was introduced into Escherichia coli BL21(DE3) (Novagen) electrocompetent cells and selected on LB agar plates containing 50 μg ml−1 carbenicillin. ..

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: Preparation of Recombinant Neutral SMase —The neutral SMase 1 cDNA was amplified by PCR using sense primer 5′-CATATGGCACCACAGCAGCCCGGCAAACTG-3′ and antisense primer 5′-CATATGTTATTCCTCCCGTTTGAAAGGACT-3′, each containing an NdeI site; the amplimer was subcloned into the pGEM-T Easy plasmid vector by the TA cloning method (Promega). .. Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1.

    Article Title: A post-translational modification signature defines changes in soluble tau correlating with oligomerization in early stage Alzheimer’s disease brain
    Article Snippet: Paragraph title: Recombinant tau protein purification ... The pET19b-Tau plasmids were transformed into E. coli BL21(DE3) cells (Novagen).

    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: Impact of enzymatic hydrolysis on the quantification of total urinary concentrations of chemical biomarkers
    Article Snippet: BL-21: β-Glucuronidase from E. coli-BL21 (Sigma Aldrich, Item #G8420, β-Glucuronidase activity ≥20,000,000 units/g protein). .. IMCS: Recombinant β-Glucuronidase from snail (Integrated Micro-Chromatography Systems Columbia, Item #04–E1F, β-glucuronidase activity > 50,000 units/mL).

    Article Title: Small Molecule Targeting TDP-43’s RNA Recognition Motifs Reduces Locomotor Defects in a Drosophila Model of Amyotrophic Lateral Sclerosis (ALS)
    Article Snippet: Paragraph title: Purification of Recombinant TDP-43 Subdomains. ... Human TDP-431–260 and TDP-43102–269 were expressed in E. coli BL21(DE3) cells (Novagen) in LB rich or M9 minimal media supplemented with 15 NH4 Cl.

    DNA Extraction:

    Article Title: Complete reconstitution of the diverse pathways of gentamicin B biosynthesis
    Article Snippet: For genomic DNA isolation and cultivation, M. echinospora was grown for 5 d at 28 °C in ATCC 172 medium (1% glucose, 0.5% yeast extract, 2% soluble starch, 0.5% N-Z amine, and 0.2% CaCO3 ). .. For expression of the recombinant proteins (GenM1, GenD2, GenS2, GenN, GenD1, GenQ, GenB1, GenJ, and GenK2), the E. coli BL21(DE3) (Novagen), E. coli BL21(DE3)pLysS (Novagen), E. coli Rosetta gamiTM2 (DE3) (Novagen) and E. coli ArcticExpress(DE3) (Agilent technologies, Santa Clara, CA, USA) were used as heterologous hosts.

    Nucleic Acid Electrophoresis:

    Article Title: Functional Characterization by Genetic Complementation of aroB-Encoded Dehydroquinate Synthase from Mycobacterium tuberculosis H37Rv and Its Heterologous Expression and Purification ▿
    Article Snippet: The recombinant pET23a(+):: aroB plasmid was introduced into Escherichia coli BL21(DE3) (Novagen) electrocompetent cells and selected on LB agar plates containing 50 μg ml−1 carbenicillin. .. Soluble and insoluble fractions were analyzed by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) ( ).

    Article Title: Molecular characterization and expression analysis of iron superoxide dismutase gene from Pseudochlorella pringsheimii (Trebouxiophyceae, Chlorophyta)
    Article Snippet: The FeSOD nucleotide sequences in the expression vector were sequenced again for validation, and then the vector was transformed into E. coli BL21(DE3)pLysS cells (Novagen, USA). .. After the E. coli cells reached a cell-density of OD600 = 0.6, the expression of the FeSOD protein was induced by the addition of 25 μM IPTG (isopropyl-β-D-thiogalactopyranoside), followed by incubation at 28 °C for 15 h. Purification of the expressed protein was done via affinity chromatography on a HiTrap HP column (GE Healthcare, USA), quantified by A280 and Bradford ( ) method (Biorad, USA) and the purity was validated by 12% SDS-PAGE (SDS–polyacrylamide gel electrophoresis; Laemmli , Fig. b).

    Subcloning:

    Article Title: Complete reconstitution of the diverse pathways of gentamicin B biosynthesis
    Article Snippet: E. coli DH5α and plasmid pGEM-T Easy Vector (Promega, Madison, WI, USA) were used for routine subcloning. .. For expression of the recombinant proteins (GenM1, GenD2, GenS2, GenN, GenD1, GenQ, GenB1, GenJ, and GenK2), the E. coli BL21(DE3) (Novagen), E. coli BL21(DE3)pLysS (Novagen), E. coli Rosetta gamiTM2 (DE3) (Novagen) and E. coli ArcticExpress(DE3) (Agilent technologies, Santa Clara, CA, USA) were used as heterologous hosts.

    Purification:

    Article Title: Functional Characterization by Genetic Complementation of aroB-Encoded Dehydroquinate Synthase from Mycobacterium tuberculosis H37Rv and Its Heterologous Expression and Purification ▿
    Article Snippet: The PCR product (1,089 bp) was purified by electrophoresis, digested with NdeI and BamHI (Boehringer Mannheim), and ligated into a pET23a(+) expression vector (Novagen) that had previously been digested with the same restriction enzymes. .. The recombinant pET23a(+):: aroB plasmid was introduced into Escherichia coli BL21(DE3) (Novagen) electrocompetent cells and selected on LB agar plates containing 50 μg ml−1 carbenicillin.

    Article Title: Cofactor-induced reversible folding of Flavodoxin-4 from Lactobacillus acidophilus
    Article Snippet: Paragraph title: Protein expression and purification ... Expression was performed in E. Coli BL21(DE3) cells (EMD Biosciences).

    Article Title: Molecular characterization and expression analysis of iron superoxide dismutase gene from Pseudochlorella pringsheimii (Trebouxiophyceae, Chlorophyta)
    Article Snippet: The FeSOD nucleotide sequences in the expression vector were sequenced again for validation, and then the vector was transformed into E. coli BL21(DE3)pLysS cells (Novagen, USA). .. After the E. coli cells reached a cell-density of OD600 = 0.6, the expression of the FeSOD protein was induced by the addition of 25 μM IPTG (isopropyl-β-D-thiogalactopyranoside), followed by incubation at 28 °C for 15 h. Purification of the expressed protein was done via affinity chromatography on a HiTrap HP column (GE Healthcare, USA), quantified by A280 and Bradford ( ) method (Biorad, USA) and the purity was validated by 12% SDS-PAGE (SDS–polyacrylamide gel electrophoresis; Laemmli , Fig. b).

    Article Title: Effects of allelic variations in the human myxovirus resistance protein A on its antiviral activity
    Article Snippet: Paragraph title: Protein expression and purification ... Human MxA and the indicated mutants/variants were expressed as N-terminal His6 -tagged fusions followed by a PreScissionTM cleavage site from a pET28 plasmid in the E. coli BL21 ( DE3 ) Rosetta strain (Novagen) as described by Gao et al . ( ).

    Article Title: The RNase YbeY Is Vital for Ribosome Maturation, Stress Resistance, and Virulence of the Natural Genetic Engineer Agrobacterium tumefaciens
    Article Snippet: Hfq6×His and YbeY6×His fusion proteins were expressed in E. coli BL21(DE3) cells, using pET-plasmid-based expression from a T7 promoter (Novagen, Madison, WI). .. After Ni-NTA chromatography, purified His-tagged fusion proteins were separated by size via a Superdex 200 10/300 GL column (HEPES buffer [pH 7.5]) on an ÄKTA system (GE Healthcare).

    Article Title: Transferrin Receptor 1-Associated Iron Accumulation and Oxidative Stress Provides a Way for Grass Carp to Fight against Reovirus Infection
    Article Snippet: To address this, pGEX-4T-1-TfR1 and pET32a(+)-Tf plasmids were transformed into the Escherichia coli BL21 and BL21(DE3) cells (Novagen), respectively. .. Then, CiTfR1 and CiTf proteins were expressed in vitro and purified.

    Article Title: Generation of a recombinant Aggregatibacter actinomycetemcomitans RTX toxin in Escherichia coli
    Article Snippet: Paragraph title: His-tagged LtxA expression and purification ... Plasmids were transformed into E. coli BL21(λDE3) cells (Novagen, Madison, WI).

    Article Title: Small Molecule Targeting TDP-43’s RNA Recognition Motifs Reduces Locomotor Defects in a Drosophila Model of Amyotrophic Lateral Sclerosis (ALS)
    Article Snippet: Paragraph title: Purification of Recombinant TDP-43 Subdomains. ... Human TDP-431–260 and TDP-43102–269 were expressed in E. coli BL21(DE3) cells (Novagen) in LB rich or M9 minimal media supplemented with 15 NH4 Cl.

    Protein Purification:

    Article Title: The RNase YbeY Is Vital for Ribosome Maturation, Stress Resistance, and Virulence of the Natural Genetic Engineer Agrobacterium tumefaciens
    Article Snippet: Paragraph title: Protein purification and GPC. ... Hfq6×His and YbeY6×His fusion proteins were expressed in E. coli BL21(DE3) cells, using pET-plasmid-based expression from a T7 promoter (Novagen, Madison, WI).

    Article Title: A post-translational modification signature defines changes in soluble tau correlating with oligomerization in early stage Alzheimer’s disease brain
    Article Snippet: Paragraph title: Recombinant tau protein purification ... The pET19b-Tau plasmids were transformed into E. coli BL21(DE3) cells (Novagen).

    Sequencing:

    Article Title: Functional Characterization by Genetic Complementation of aroB-Encoded Dehydroquinate Synthase from Mycobacterium tuberculosis H37Rv and Its Heterologous Expression and Purification ▿
    Article Snippet: The DNA sequence of the M. tuberculosis aroB gene was determined in order to confirm the identity, integrity, and absence of PCR-introduced mutations in the cloned gene. .. The recombinant pET23a(+):: aroB plasmid was introduced into Escherichia coli BL21(DE3) (Novagen) electrocompetent cells and selected on LB agar plates containing 50 μg ml−1 carbenicillin.

    Article Title: Cofactor-induced reversible folding of Flavodoxin-4 from Lactobacillus acidophilus
    Article Snippet: The LBA1001 gene (Uniprot ID: Q5FKC3, GenBank: ) was amplified by polymerase chain reaction (PCR) from Lactobacillus acidophilus genomic DNA using PfuTurbo DNA polymerase (Stratagene) and I-PIPE (forward primer, 5′-ctgtacttccagggc ATGGCTAAAAAAACATTAATTTTAT-3′; reverse primer, 5′-aattaagtcgcgtta TTTACTCCATTTATCAACTTCACTATC-3′, target sequence in upper case) that included sequences for the predicted 5′ and 3′ ends. .. Expression was performed in E. Coli BL21(DE3) cells (EMD Biosciences).

    Article Title: Molecular characterization and expression analysis of iron superoxide dismutase gene from Pseudochlorella pringsheimii (Trebouxiophyceae, Chlorophyta)
    Article Snippet: The FeSOD nucleotide sequences in the expression vector were sequenced again for validation, and then the vector was transformed into E. coli BL21(DE3)pLysS cells (Novagen, USA). .. The nucleotide sequence of the FeSOD of P. pringsheimii was deposited at the NCBI GenBank database under accession number .

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: The cloned nucleotide sequence was confirmed by sequencing and subcloned into the NdeI site in the multiple cloning site of the pET-16b vector (Novagen) to fuse a His10 tag sequence to the N terminus of the neutral SMase 1 open reading frame. .. Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1.

    Article Title: Dystrophin As A Molecular Shock Absorber
    Article Snippet: Four fragments of the dystrophin central rod domain ( i.e ., SR01–05: 338th -938th ; SR06–10: 939th -1466th ; SR11–17: 1464th -2210th ; SR18–24: 2209th -3044th ) were synthesized by PCR using plasmid template containing the full-length human dystrophin sequence. .. Each of the resulting plasmids was co-transformed with a BirA plasmid and expressed in Escherichia coli BL21 (DE3) cultured in LB-media with D-Biotin (Sigma Aldrich), and affinity purified through His-tag.

    Positron Emission Tomography:

    Article Title: Complete reconstitution of the diverse pathways of gentamicin B biosynthesis
    Article Snippet: The pET expression vectors, such as pET15b and pET28a, were purchased from Novagen (Madison, WI, USA). .. For expression of the recombinant proteins (GenM1, GenD2, GenS2, GenN, GenD1, GenQ, GenB1, GenJ, and GenK2), the E. coli BL21(DE3) (Novagen), E. coli BL21(DE3)pLysS (Novagen), E. coli Rosetta gamiTM2 (DE3) (Novagen) and E. coli ArcticExpress(DE3) (Agilent technologies, Santa Clara, CA, USA) were used as heterologous hosts.

    Article Title: The RNase YbeY Is Vital for Ribosome Maturation, Stress Resistance, and Virulence of the Natural Genetic Engineer Agrobacterium tumefaciens
    Article Snippet: .. Hfq6×His and YbeY6×His fusion proteins were expressed in E. coli BL21(DE3) cells, using pET-plasmid-based expression from a T7 promoter (Novagen, Madison, WI). .. To assess the oligomeric state, we performed gel permeation chromatography.

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: The cloned nucleotide sequence was confirmed by sequencing and subcloned into the NdeI site in the multiple cloning site of the pET-16b vector (Novagen) to fuse a His10 tag sequence to the N terminus of the neutral SMase 1 open reading frame. .. Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1.

    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.

    SDS Page:

    Article Title: Functional Characterization by Genetic Complementation of aroB-Encoded Dehydroquinate Synthase from Mycobacterium tuberculosis H37Rv and Its Heterologous Expression and Purification ▿
    Article Snippet: The recombinant pET23a(+):: aroB plasmid was introduced into Escherichia coli BL21(DE3) (Novagen) electrocompetent cells and selected on LB agar plates containing 50 μg ml−1 carbenicillin. .. Soluble and insoluble fractions were analyzed by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) ( ).

    Article Title: Molecular characterization and expression analysis of iron superoxide dismutase gene from Pseudochlorella pringsheimii (Trebouxiophyceae, Chlorophyta)
    Article Snippet: The FeSOD nucleotide sequences in the expression vector were sequenced again for validation, and then the vector was transformed into E. coli BL21(DE3)pLysS cells (Novagen, USA). .. After the E. coli cells reached a cell-density of OD600 = 0.6, the expression of the FeSOD protein was induced by the addition of 25 μM IPTG (isopropyl-β-D-thiogalactopyranoside), followed by incubation at 28 °C for 15 h. Purification of the expressed protein was done via affinity chromatography on a HiTrap HP column (GE Healthcare, USA), quantified by A280 and Bradford ( ) method (Biorad, USA) and the purity was validated by 12% SDS-PAGE (SDS–polyacrylamide gel electrophoresis; Laemmli , Fig. b).

    Plasmid Preparation:

    Article Title: Complete reconstitution of the diverse pathways of gentamicin B biosynthesis
    Article Snippet: E. coli DH5α and plasmid pGEM-T Easy Vector (Promega, Madison, WI, USA) were used for routine subcloning. .. For expression of the recombinant proteins (GenM1, GenD2, GenS2, GenN, GenD1, GenQ, GenB1, GenJ, and GenK2), the E. coli BL21(DE3) (Novagen), E. coli BL21(DE3)pLysS (Novagen), E. coli Rosetta gamiTM2 (DE3) (Novagen) and E. coli ArcticExpress(DE3) (Agilent technologies, Santa Clara, CA, USA) were used as heterologous hosts.

    Article Title: Functional Characterization by Genetic Complementation of aroB-Encoded Dehydroquinate Synthase from Mycobacterium tuberculosis H37Rv and Its Heterologous Expression and Purification ▿
    Article Snippet: .. The recombinant pET23a(+):: aroB plasmid was introduced into Escherichia coli BL21(DE3) (Novagen) electrocompetent cells and selected on LB agar plates containing 50 μg ml−1 carbenicillin. ..

    Article Title: Cofactor-induced reversible folding of Flavodoxin-4 from Lactobacillus acidophilus
    Article Snippet: The expression vector, pSpeedET, which encodes an amino-terminal tobacco etch virus (TEV) protease-cleavable expression and purification tag (MGSDKIHHHHHHENLYFQ-G), was PCR amplified with V-PIPE (Vector) primers (forward primer: 5′-taacgcgacttaattaactcgtttaaacggtctccagc-3′, reverse primer: 5′-gccctggaagtacaggttttcgtgatgatgatgatgatg-3′). .. Expression was performed in E. Coli BL21(DE3) cells (EMD Biosciences).

    Article Title: Molecular characterization and expression analysis of iron superoxide dismutase gene from Pseudochlorella pringsheimii (Trebouxiophyceae, Chlorophyta)
    Article Snippet: .. The FeSOD nucleotide sequences in the expression vector were sequenced again for validation, and then the vector was transformed into E. coli BL21(DE3)pLysS cells (Novagen, USA). .. After the E. coli cells reached a cell-density of OD600 = 0.6, the expression of the FeSOD protein was induced by the addition of 25 μM IPTG (isopropyl-β-D-thiogalactopyranoside), followed by incubation at 28 °C for 15 h. Purification of the expressed protein was done via affinity chromatography on a HiTrap HP column (GE Healthcare, USA), quantified by A280 and Bradford ( ) method (Biorad, USA) and the purity was validated by 12% SDS-PAGE (SDS–polyacrylamide gel electrophoresis; Laemmli , Fig. b).

    Article Title: Effects of allelic variations in the human myxovirus resistance protein A on its antiviral activity
    Article Snippet: .. Human MxA and the indicated mutants/variants were expressed as N-terminal His6 -tagged fusions followed by a PreScissionTM cleavage site from a pET28 plasmid in the E. coli BL21 ( DE3 ) Rosetta strain (Novagen) as described by Gao et al . ( ). ..

    Article Title: A Novel Prokaryote-Type ECF/ABC Transporter Module in Chloroplast Metal Homeostasis
    Article Snippet: .. The resulting mature versions of At-ABCI10 and At-ABCI11 were subcloned into the pET21d (Novagen) plasmid vector and used for overexpression after transforming E. coli BL21(DE3) cells (Novagen). ..

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: The cloned nucleotide sequence was confirmed by sequencing and subcloned into the NdeI site in the multiple cloning site of the pET-16b vector (Novagen) to fuse a His10 tag sequence to the N terminus of the neutral SMase 1 open reading frame. .. Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1.

    Article Title: A post-translational modification signature defines changes in soluble tau correlating with oligomerization in early stage Alzheimer’s disease brain
    Article Snippet: Recombinant tau protein purification Tau variants (full length protein and a fragment encoding amino acids 256–368) were cloned into the pET19b vector (Novagen) in between the NcoI and BamHI restriction sites. .. The pET19b-Tau plasmids were transformed into E. coli BL21(DE3) cells (Novagen).

    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: Dystrophin As A Molecular Shock Absorber
    Article Snippet: .. Each of the resulting plasmids was co-transformed with a BirA plasmid and expressed in Escherichia coli BL21 (DE3) cultured in LB-media with D-Biotin (Sigma Aldrich), and affinity purified through His-tag. .. A vertical magnetic tweezers setup, , was combined with a disturbance-free, rapid solution-exchange flow channel for conducting in vitro protein stretching experiments.

    Affinity Chromatography:

    Article Title: Molecular characterization and expression analysis of iron superoxide dismutase gene from Pseudochlorella pringsheimii (Trebouxiophyceae, Chlorophyta)
    Article Snippet: The FeSOD nucleotide sequences in the expression vector were sequenced again for validation, and then the vector was transformed into E. coli BL21(DE3)pLysS cells (Novagen, USA). .. After the E. coli cells reached a cell-density of OD600 = 0.6, the expression of the FeSOD protein was induced by the addition of 25 μM IPTG (isopropyl-β-D-thiogalactopyranoside), followed by incubation at 28 °C for 15 h. Purification of the expressed protein was done via affinity chromatography on a HiTrap HP column (GE Healthcare, USA), quantified by A280 and Bradford ( ) method (Biorad, USA) and the purity was validated by 12% SDS-PAGE (SDS–polyacrylamide gel electrophoresis; Laemmli , Fig. b).

    In Vitro:

    Article Title: Transferrin Receptor 1-Associated Iron Accumulation and Oxidative Stress Provides a Way for Grass Carp to Fight against Reovirus Infection
    Article Snippet: To address this, pGEX-4T-1-TfR1 and pET32a(+)-Tf plasmids were transformed into the Escherichia coli BL21 and BL21(DE3) cells (Novagen), respectively. .. Then, CiTfR1 and CiTf proteins were expressed in vitro and purified.

    Gel Permeation Chromatography:

    Article Title: The RNase YbeY Is Vital for Ribosome Maturation, Stress Resistance, and Virulence of the Natural Genetic Engineer Agrobacterium tumefaciens
    Article Snippet: Paragraph title: Protein purification and GPC. ... Hfq6×His and YbeY6×His fusion proteins were expressed in E. coli BL21(DE3) cells, using pET-plasmid-based expression from a T7 promoter (Novagen, Madison, WI).

    Concentration Assay:

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1. .. Isopropyl-1-thio-β- d -galactopyranoside was added to a final concentration of 1 m m , and the culture was grown for an additional 4 h to induce the expression of the transgene.

    Marker:

    Article Title: Complete reconstitution of the diverse pathways of gentamicin B biosynthesis
    Article Snippet: For expression of the recombinant proteins (GenM1, GenD2, GenS2, GenN, GenD1, GenQ, GenB1, GenJ, and GenK2), the E. coli BL21(DE3) (Novagen), E. coli BL21(DE3)pLysS (Novagen), E. coli Rosetta gamiTM2 (DE3) (Novagen) and E. coli ArcticExpress(DE3) (Agilent technologies, Santa Clara, CA, USA) were used as heterologous hosts. .. For the recombinant GenM2, the high-copy-number E. coli -Streptomyces shuttle vector pSE34 containing the constitutive ermE * promoter plus a thiostrepton resistance marker was used for expression .

    Lysis:

    Article Title: A Novel Prokaryote-Type ECF/ABC Transporter Module in Chloroplast Metal Homeostasis
    Article Snippet: The resulting mature versions of At-ABCI10 and At-ABCI11 were subcloned into the pET21d (Novagen) plasmid vector and used for overexpression after transforming E. coli BL21(DE3) cells (Novagen). .. Afterwards, pelleted cells (4°C, 6,000g, 15 min) were resuspended in cell lysis solution (50 mM Tris-HCl, pH 8.0, 25% [w/v] sucrose, 1 mM EDTA, 100 mg/ml DNase) and sonicated three times for 30 s. Inclusion bodies were collected by centrifugation at 4°C and 20,000g for 30 min.

    Article Title: Identification of Mg2+-dependent Neutral Sphingomyelinase 1 as a Mediator of Heat Stress-induced Ceramide Generation and Apoptosis *
    Article Snippet: At 24 h after transfection, the cells were washed twice with PBS and homogenized in lysis buffer for neutral SMase assay. .. Neutral SMase 1 was expressed in E. coli BL21(DE3)pLysE cells (Novagen) transformed with pETZNSMase 1.

    Activity Assay:

    Article Title: Impact of enzymatic hydrolysis on the quantification of total urinary concentrations of chemical biomarkers
    Article Snippet: .. BL-21: β-Glucuronidase from E. coli-BL21 (Sigma Aldrich, Item #G8420, β-Glucuronidase activity ≥20,000,000 units/g protein). .. HP-2: β-Glucuronidase from Helix Pomatia, Type HP-2 (Sigma Aldrich, Item #G7017, β-glucuronidase activity ≥ 100,000 units/mL; sulfatase activity 7500 units/mL).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    Millipore e coli bl21
    E Coli Bl21, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 865 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli bl21/product/Millipore
    Average 90 stars, based on 865 article reviews
    Price from $9.99 to $1999.99
    e coli bl21 - by Bioz Stars, 2020-01
    90/100 stars
      Buy from Supplier

    Image Search Results