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Merck KGaA escherichia coli strain bl21
Escherichia Coli Strain Bl21, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Clone Assay:

Article Title: Solubilization and renaturation of biologically active human bone morphogenetic protein-4 from inclusion bodies
Article Snippet: The cloning procedure was verified by DNA sequencing. .. The expression vector pET24a-hBMP4 was then transformed into the E. coli strain BL21(DE3) (Merck Millipore, USA).

Article Title: De novo GTP Biosynthesis Is Critical for Virulence of the Fungal Pathogen Cryptococcus neoformans
Article Snippet: Cloning and plasmid propagation was performed in E. coli strain Mach1 (Life Technologies, Carlsbad CA). .. Recombinant Cryptococcus IMPDH was heterologously expressed in E. coli strain BL21(DE3)pLysS (Merck, Darmstadt, Germany).

Centrifugation:

Article Title: Truncated Hantavirus Nucleocapsid Proteins for Serotyping Sin Nombre, Andes, and Laguna Negra Hantavirus Infections in Humans and Rodents ▿
Article Snippet: The DNA fractions were subcloned into the pET43b vector (Merck KGaA, Darmstadt, Germany) and transfected into E. coli strain BL21(DE3) (Merck KGaA). .. The cultured cells were collected by centrifugation, resuspended in 5 ml of 0.5 M NaCl binding buffer (0.5 M NaCl, 20 mM imidazole, 20 mM potassium phosphate), and sonicated on ice four times for 15 s each time.

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: E. coli strain BL21 (Merck Millipore), transformed with pCold-rhaR, was grown with shaking in a Luria-Bertani medium ( ) supplemented with ampicillin (50 μg/ml) at 37°C to an OD600 of 0.4. .. After centrifugation (17,000 × g , 4°C, 20 min) and filtration (0.45-μm pore size), the supernatant was subjected to ammonium sulfate precipitation.

Article Title: The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis
Article Snippet: The recombinant protein with an N-terminal His6 -tag was produced in Escherichia coli strain BL21(DE3) (Merck, Germany) at 22 °C with an incubation for 16–20 h after induction with 0.5 mM isopropyl β-d -1-thiogalactopyranoside (IPTG). .. Bacteria were lysed by sonication on ice at 200 W using 3-s pulses with 7-s intervals for 16.5 min before the removal of insoluble debris by centrifugation for 30 min at 13,000 × g and 4 °C.

Amplification:

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: The B. subtilis rhaR , rhaA , and rhaB coding regions were amplified by PCR with genomic DNA of strain 168 and the primer pairs rhaRORF_NF/rhaRORF_BR, rhaAORF_XF/rhaAORF_BR, and rhaBORF_XF/rhaBORF_BR, respectively, and the E. coli rhaB coding region was amplified by PCR with genomic DNA of strain DH5α and the primer pair EcrhaB_XF/EcrhaB_BR ( ). .. E. coli strain BL21 (Merck Millipore), transformed with pCold-rhaR, was grown with shaking in a Luria-Bertani medium ( ) supplemented with ampicillin (50 μg/ml) at 37°C to an OD600 of 0.4.

Article Title: The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis
Article Snippet: The amplified product was later inserted into a pET28a vector between Nde I and Xho I restriction sites. .. The recombinant protein with an N-terminal His6 -tag was produced in Escherichia coli strain BL21(DE3) (Merck, Germany) at 22 °C with an incubation for 16–20 h after induction with 0.5 mM isopropyl β-d -1-thiogalactopyranoside (IPTG).

Filtration:

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: E. coli strain BL21 (Merck Millipore), transformed with pCold-rhaR, was grown with shaking in a Luria-Bertani medium ( ) supplemented with ampicillin (50 μg/ml) at 37°C to an OD600 of 0.4. .. After centrifugation (17,000 × g , 4°C, 20 min) and filtration (0.45-μm pore size), the supernatant was subjected to ammonium sulfate precipitation.

Synthesized:

Article Title: Solubilization and renaturation of biologically active human bone morphogenetic protein-4 from inclusion bodies
Article Snippet: The gene coding for hBMP-4 was synthesized as a codon-optimized sequence for E. coli and cloned into the expression vector pET24a. .. The expression vector pET24a-hBMP4 was then transformed into the E. coli strain BL21(DE3) (Merck Millipore, USA).

Article Title: Solubilization and renaturation of biologically active human bone morphogenetic protein-4 from inclusion bodies
Article Snippet: 2.1 Organism and growth conditions The gene coding for hBMP-4 was synthesized as a codon-optimized sequence for E. coli and cloned into the expression vector pET24a. .. The expression vector pET24a-hBMP4 was then transformed into the E. coli strain BL21(DE3) (Merck Millipore, USA).

Article Title: The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis
Article Snippet: Protein expression and purification A coding sequence for full-length NDM-1 (Supplementary Table ) was synthesized using GENEWIZ (Sangon Biotech, Shanghai, China). .. The recombinant protein with an N-terminal His6 -tag was produced in Escherichia coli strain BL21(DE3) (Merck, Germany) at 22 °C with an incubation for 16–20 h after induction with 0.5 mM isopropyl β-d -1-thiogalactopyranoside (IPTG).

Incubation:

Article Title: Truncated Hantavirus Nucleocapsid Proteins for Serotyping Sin Nombre, Andes, and Laguna Negra Hantavirus Infections in Humans and Rodents ▿
Article Snippet: The DNA fractions were subcloned into the pET43b vector (Merck KGaA, Darmstadt, Germany) and transfected into E. coli strain BL21(DE3) (Merck KGaA). .. A single colony was inoculated into Circle growth medium (MP Biomedicals, Morgan Irvine, CA) containing ampicillin (50 μg/ml) for small-scale culture incubation at 37°C overnight.

Article Title: The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis
Article Snippet: .. The recombinant protein with an N-terminal His6 -tag was produced in Escherichia coli strain BL21(DE3) (Merck, Germany) at 22 °C with an incubation for 16–20 h after induction with 0.5 mM isopropyl β-d -1-thiogalactopyranoside (IPTG). .. Harvested cells were resuspended in the lysis buffer containing 50 mM Tris pH 8.0, 500 mM NaCl, 10 mM imidazole, 10 mM β-mercaptoethanol, and 1 mM ZnCl2 .

Expressing:

Article Title: Truncated Hantavirus Nucleocapsid Proteins for Serotyping Sin Nombre, Andes, and Laguna Negra Hantavirus Infections in Humans and Rodents ▿
Article Snippet: The DNA fractions were subcloned into the pET43b vector (Merck KGaA, Darmstadt, Germany) and transfected into E. coli strain BL21(DE3) (Merck KGaA). .. The culture fluid was then centrifuged, the collected cells were inoculated into 100 ml of Circle growth medium, and isopropyl-β- d -1-thiogalactopyranoside (IPTG) induction was performed according to the procedure for pET system expression.

Article Title: Efficient Single-Strand Break Repair Requires Binding to Both Poly(ADP-Ribose) and DNA by the Central BRCT Domain of XRCC1
Article Snippet: Paragraph title: Expression and purification ... E. coli strain BL21(DE3) (Merck Millipore) was co-transformed with pET15b-SUMO-XRCC1-BRCT1 plasmid.

Article Title: Solubilization and renaturation of biologically active human bone morphogenetic protein-4 from inclusion bodies
Article Snippet: .. The expression vector pET24a-hBMP4 was then transformed into the E. coli strain BL21(DE3) (Merck Millipore, USA). ..

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: The other three fragments were digested with XhoI/BamHI and then cloned into the pColdI vector (TaKaRa Bio), which had been treated with the same restriction enzymes, to yield expression plasmids pCold-rhaA, pCold-rhaB, and pCold-EcrhaB. .. E. coli strain BL21 (Merck Millipore), transformed with pCold-rhaR, was grown with shaking in a Luria-Bertani medium ( ) supplemented with ampicillin (50 μg/ml) at 37°C to an OD600 of 0.4.

Article Title: Phosphorylation of serine residues in the N-terminus modulates the activity of ACA8, a plasma membrane Ca2+-ATPase of Arabidopsis thaliana
Article Snippet: .. Expression and purification of the WT and mutated His-tagged ACA8 N-terminus Vectors coding for WT and mutated His-tagged ACA8 N-terminus (6His-1 M-I116 ) were used to transform E. coli strain BL21(DE3)pLysE (Merck KGaA, Darmstadt, Germany, catalogue no. 69389-3) by standard procedures. ..

Article Title: Solubilization and renaturation of biologically active human bone morphogenetic protein-4 from inclusion bodies
Article Snippet: .. The expression vector pET24a-hBMP4 was then transformed into the E. coli strain BL21(DE3) (Merck Millipore, USA). ..

Article Title: The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis
Article Snippet: Paragraph title: Protein expression and purification ... The recombinant protein with an N-terminal His6 -tag was produced in Escherichia coli strain BL21(DE3) (Merck, Germany) at 22 °C with an incubation for 16–20 h after induction with 0.5 mM isopropyl β-d -1-thiogalactopyranoside (IPTG).

Transformation Assay:

Article Title: Solubilization and renaturation of biologically active human bone morphogenetic protein-4 from inclusion bodies
Article Snippet: .. The expression vector pET24a-hBMP4 was then transformed into the E. coli strain BL21(DE3) (Merck Millipore, USA). ..

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: .. E. coli strain BL21 (Merck Millipore), transformed with pCold-rhaR, was grown with shaking in a Luria-Bertani medium ( ) supplemented with ampicillin (50 μg/ml) at 37°C to an OD600 of 0.4. .. The culture was then refrigerated at 15°C and left to stand for 30 min. After IPTG had been added to a final concentration of 1 mM, shaking of the culture was resumed at 15°C and continued for another 24 h. The cells harvested from 6 liters of the culture were disrupted by sonication in buffer 1 (20 mM Tris-Cl buffer [pH 8.0] plus 10% [vol/vol] glycerol, 0.1 mM phenylmethylsulfonyl fluoride, and 1 mM dithiothreitol).

Article Title: Solubilization and renaturation of biologically active human bone morphogenetic protein-4 from inclusion bodies
Article Snippet: .. The expression vector pET24a-hBMP4 was then transformed into the E. coli strain BL21(DE3) (Merck Millipore, USA). ..

Transfection:

Article Title: Truncated Hantavirus Nucleocapsid Proteins for Serotyping Sin Nombre, Andes, and Laguna Negra Hantavirus Infections in Humans and Rodents ▿
Article Snippet: .. The DNA fractions were subcloned into the pET43b vector (Merck KGaA, Darmstadt, Germany) and transfected into E. coli strain BL21(DE3) (Merck KGaA). .. A single colony was inoculated into Circle growth medium (MP Biomedicals, Morgan Irvine, CA) containing ampicillin (50 μg/ml) for small-scale culture incubation at 37°C overnight.

Chromatography:

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: The HPr protein, produced by E. coli strain BL21(DE3) (Merck Millipore) bearing plasmid pET-ptsH, was purified by one column chromatography step (Toyopearl DEAE-650M). .. An N-terminal in-frame fusion of B. subtilis HPr kinase/phosphorylase with the His6 tag (His-HPrK), produced by E. coli strain BL21(DE3) bearing plasmid pET-hprK, was purified by affinity column chromatography (Ni-nitrilotriacetic acid [NTA]-agarose; Qiagen).

Article Title: The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis
Article Snippet: The recombinant protein with an N-terminal His6 -tag was produced in Escherichia coli strain BL21(DE3) (Merck, Germany) at 22 °C with an incubation for 16–20 h after induction with 0.5 mM isopropyl β-d -1-thiogalactopyranoside (IPTG). .. The supernatant was immediately loaded onto a Ni2+ -NTA chromatography column (Novagen), followed by column washing and elution with 250 mM imidazole added in the same buffer.

Concentration Assay:

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: E. coli strain BL21 (Merck Millipore), transformed with pCold-rhaR, was grown with shaking in a Luria-Bertani medium ( ) supplemented with ampicillin (50 μg/ml) at 37°C to an OD600 of 0.4. .. The culture was then refrigerated at 15°C and left to stand for 30 min. After IPTG had been added to a final concentration of 1 mM, shaking of the culture was resumed at 15°C and continued for another 24 h. The cells harvested from 6 liters of the culture were disrupted by sonication in buffer 1 (20 mM Tris-Cl buffer [pH 8.0] plus 10% [vol/vol] glycerol, 0.1 mM phenylmethylsulfonyl fluoride, and 1 mM dithiothreitol).

Article Title: De novo GTP Biosynthesis Is Critical for Virulence of the Fungal Pathogen Cryptococcus neoformans
Article Snippet: Recombinant Cryptococcus IMPDH was heterologously expressed in E. coli strain BL21(DE3)pLysS (Merck, Darmstadt, Germany). .. MPA was dissolved in DMSO and was used at a concentration of 5 µg/mL.

Cell Culture:

Article Title: Truncated Hantavirus Nucleocapsid Proteins for Serotyping Sin Nombre, Andes, and Laguna Negra Hantavirus Infections in Humans and Rodents ▿
Article Snippet: The DNA fractions were subcloned into the pET43b vector (Merck KGaA, Darmstadt, Germany) and transfected into E. coli strain BL21(DE3) (Merck KGaA). .. The cultured cells were collected by centrifugation, resuspended in 5 ml of 0.5 M NaCl binding buffer (0.5 M NaCl, 20 mM imidazole, 20 mM potassium phosphate), and sonicated on ice four times for 15 s each time.

Article Title: De novo GTP Biosynthesis Is Critical for Virulence of the Fungal Pathogen Cryptococcus neoformans
Article Snippet: Strains and media C. neoformans and C. gattii strains were cultured in YPD or YNB (Becton Dickinson, Franklin Lakes NJ) media at 30°C. imd1Δ mutants are guanine auxotrophs and were grown on minimal YNB media supplemented with 1 mM guanine for all manipulations; ade2Δ and hpt1Δ ade2Δ mutants are adenine auxotrophs and were maintained on rich media. .. Recombinant Cryptococcus IMPDH was heterologously expressed in E. coli strain BL21(DE3)pLysS (Merck, Darmstadt, Germany).

DNA Sequencing:

Article Title: Solubilization and renaturation of biologically active human bone morphogenetic protein-4 from inclusion bodies
Article Snippet: The cloning procedure was verified by DNA sequencing. .. The expression vector pET24a-hBMP4 was then transformed into the E. coli strain BL21(DE3) (Merck Millipore, USA).

Sequencing:

Article Title: Solubilization and renaturation of biologically active human bone morphogenetic protein-4 from inclusion bodies
Article Snippet: The gene coding for hBMP-4 was synthesized as a codon-optimized sequence for E. coli and cloned into the expression vector pET24a. .. The expression vector pET24a-hBMP4 was then transformed into the E. coli strain BL21(DE3) (Merck Millipore, USA).

Article Title: Solubilization and renaturation of biologically active human bone morphogenetic protein-4 from inclusion bodies
Article Snippet: 2.1 Organism and growth conditions The gene coding for hBMP-4 was synthesized as a codon-optimized sequence for E. coli and cloned into the expression vector pET24a. .. The expression vector pET24a-hBMP4 was then transformed into the E. coli strain BL21(DE3) (Merck Millipore, USA).

Article Title: The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis
Article Snippet: The nucleotide sequence encoding an N terminus truncated protein (residues 29–270) was amplified with a pair of primers (Supplementary Table ) by PCR. .. The recombinant protein with an N-terminal His6 -tag was produced in Escherichia coli strain BL21(DE3) (Merck, Germany) at 22 °C with an incubation for 16–20 h after induction with 0.5 mM isopropyl β-d -1-thiogalactopyranoside (IPTG).

Sonication:

Article Title: Truncated Hantavirus Nucleocapsid Proteins for Serotyping Sin Nombre, Andes, and Laguna Negra Hantavirus Infections in Humans and Rodents ▿
Article Snippet: The DNA fractions were subcloned into the pET43b vector (Merck KGaA, Darmstadt, Germany) and transfected into E. coli strain BL21(DE3) (Merck KGaA). .. The cultured cells were collected by centrifugation, resuspended in 5 ml of 0.5 M NaCl binding buffer (0.5 M NaCl, 20 mM imidazole, 20 mM potassium phosphate), and sonicated on ice four times for 15 s each time.

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: E. coli strain BL21 (Merck Millipore), transformed with pCold-rhaR, was grown with shaking in a Luria-Bertani medium ( ) supplemented with ampicillin (50 μg/ml) at 37°C to an OD600 of 0.4. .. The culture was then refrigerated at 15°C and left to stand for 30 min. After IPTG had been added to a final concentration of 1 mM, shaking of the culture was resumed at 15°C and continued for another 24 h. The cells harvested from 6 liters of the culture were disrupted by sonication in buffer 1 (20 mM Tris-Cl buffer [pH 8.0] plus 10% [vol/vol] glycerol, 0.1 mM phenylmethylsulfonyl fluoride, and 1 mM dithiothreitol).

Article Title: The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis
Article Snippet: The recombinant protein with an N-terminal His6 -tag was produced in Escherichia coli strain BL21(DE3) (Merck, Germany) at 22 °C with an incubation for 16–20 h after induction with 0.5 mM isopropyl β-d -1-thiogalactopyranoside (IPTG). .. Bacteria were lysed by sonication on ice at 200 W using 3-s pulses with 7-s intervals for 16.5 min before the removal of insoluble debris by centrifugation for 30 min at 13,000 × g and 4 °C.

Recombinant:

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: Paragraph title: Preparation of the recombinant proteins. ... E. coli strain BL21 (Merck Millipore), transformed with pCold-rhaR, was grown with shaking in a Luria-Bertani medium ( ) supplemented with ampicillin (50 μg/ml) at 37°C to an OD600 of 0.4.

Article Title: De novo GTP Biosynthesis Is Critical for Virulence of the Fungal Pathogen Cryptococcus neoformans
Article Snippet: .. Recombinant Cryptococcus IMPDH was heterologously expressed in E. coli strain BL21(DE3)pLysS (Merck, Darmstadt, Germany). .. MPA, mizoribine, 5-fluoro-2′-deoxyuridine, G418, NAD+ , Tris, Bis-Tris, glycine, NaCl, KCl, DTT, EDTA, DMSO, uric acid, inosine, hypoxanthine, xanthine, xanthosine, guanine, guanosine, adenine, adenosine, IMP, AMP, XMP and GMP were purchased from Sigma (St Louis, MO).

Article Title: The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis
Article Snippet: .. The recombinant protein with an N-terminal His6 -tag was produced in Escherichia coli strain BL21(DE3) (Merck, Germany) at 22 °C with an incubation for 16–20 h after induction with 0.5 mM isopropyl β-d -1-thiogalactopyranoside (IPTG). .. Harvested cells were resuspended in the lysis buffer containing 50 mM Tris pH 8.0, 500 mM NaCl, 10 mM imidazole, 10 mM β-mercaptoethanol, and 1 mM ZnCl2 .

Mutagenesis:

Article Title: Histone H1 Recruitment by CHD8 Is Essential for Suppression of the Wnt-?-Catenin Signaling Pathway
Article Snippet: Complementary DNAs encoding wild-type or mutant forms of mouse CHD8S tagged with the FLAG or Myc epitope at the NH2 terminus were subcloned into pcDNA3 (Invitrogen) with the use of the Gateway vector conversion system (Invitrogen); those encoding wild-type or mutant forms of mouse histone H1c with three copies of the FLAG epitope at the NH2 terminus were also subcloned into pcDNA3. .. His6 -tagged proteins were expressed in Escherichia coli strain BL21(DE3)pLys(S) (Merck, Darmstadt, Germany).

Isolation:

Article Title: De novo GTP Biosynthesis Is Critical for Virulence of the Fungal Pathogen Cryptococcus neoformans
Article Snippet: Genomic DNA for E. coli gene isolation was prepared from strain K12. .. Recombinant Cryptococcus IMPDH was heterologously expressed in E. coli strain BL21(DE3)pLysS (Merck, Darmstadt, Germany).

Purification:

Article Title: Efficient Single-Strand Break Repair Requires Binding to Both Poly(ADP-Ribose) and DNA by the Central BRCT Domain of XRCC1
Article Snippet: Paragraph title: Expression and purification ... E. coli strain BL21(DE3) (Merck Millipore) was co-transformed with pET15b-SUMO-XRCC1-BRCT1 plasmid.

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: .. The HPr protein, produced by E. coli strain BL21(DE3) (Merck Millipore) bearing plasmid pET-ptsH, was purified by one column chromatography step (Toyopearl DEAE-650M). .. An N-terminal in-frame fusion of B. subtilis HPr kinase/phosphorylase with the His6 tag (His-HPrK), produced by E. coli strain BL21(DE3) bearing plasmid pET-hprK, was purified by affinity column chromatography (Ni-nitrilotriacetic acid [NTA]-agarose; Qiagen).

Article Title: Phosphorylation of serine residues in the N-terminus modulates the activity of ACA8, a plasma membrane Ca2+-ATPase of Arabidopsis thaliana
Article Snippet: .. Expression and purification of the WT and mutated His-tagged ACA8 N-terminus Vectors coding for WT and mutated His-tagged ACA8 N-terminus (6His-1 M-I116 ) were used to transform E. coli strain BL21(DE3)pLysE (Merck KGaA, Darmstadt, Germany, catalogue no. 69389-3) by standard procedures. ..

Article Title: The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis
Article Snippet: Paragraph title: Protein expression and purification ... The recombinant protein with an N-terminal His6 -tag was produced in Escherichia coli strain BL21(DE3) (Merck, Germany) at 22 °C with an incubation for 16–20 h after induction with 0.5 mM isopropyl β-d -1-thiogalactopyranoside (IPTG).

Polymerase Chain Reaction:

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: The B. subtilis rhaR , rhaA , and rhaB coding regions were amplified by PCR with genomic DNA of strain 168 and the primer pairs rhaRORF_NF/rhaRORF_BR, rhaAORF_XF/rhaAORF_BR, and rhaBORF_XF/rhaBORF_BR, respectively, and the E. coli rhaB coding region was amplified by PCR with genomic DNA of strain DH5α and the primer pair EcrhaB_XF/EcrhaB_BR ( ). .. E. coli strain BL21 (Merck Millipore), transformed with pCold-rhaR, was grown with shaking in a Luria-Bertani medium ( ) supplemented with ampicillin (50 μg/ml) at 37°C to an OD600 of 0.4.

Article Title: The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis
Article Snippet: The nucleotide sequence encoding an N terminus truncated protein (residues 29–270) was amplified with a pair of primers (Supplementary Table ) by PCR. .. The recombinant protein with an N-terminal His6 -tag was produced in Escherichia coli strain BL21(DE3) (Merck, Germany) at 22 °C with an incubation for 16–20 h after induction with 0.5 mM isopropyl β-d -1-thiogalactopyranoside (IPTG).

Positron Emission Tomography:

Article Title: Truncated Hantavirus Nucleocapsid Proteins for Serotyping Sin Nombre, Andes, and Laguna Negra Hantavirus Infections in Humans and Rodents ▿
Article Snippet: The DNA fractions were subcloned into the pET43b vector (Merck KGaA, Darmstadt, Germany) and transfected into E. coli strain BL21(DE3) (Merck KGaA). .. The culture fluid was then centrifuged, the collected cells were inoculated into 100 ml of Circle growth medium, and isopropyl-β- d -1-thiogalactopyranoside (IPTG) induction was performed according to the procedure for pET system expression.

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: .. The HPr protein, produced by E. coli strain BL21(DE3) (Merck Millipore) bearing plasmid pET-ptsH, was purified by one column chromatography step (Toyopearl DEAE-650M). .. An N-terminal in-frame fusion of B. subtilis HPr kinase/phosphorylase with the His6 tag (His-HPrK), produced by E. coli strain BL21(DE3) bearing plasmid pET-hprK, was purified by affinity column chromatography (Ni-nitrilotriacetic acid [NTA]-agarose; Qiagen).

Affinity Column:

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: The HPr protein, produced by E. coli strain BL21(DE3) (Merck Millipore) bearing plasmid pET-ptsH, was purified by one column chromatography step (Toyopearl DEAE-650M). .. An N-terminal in-frame fusion of B. subtilis HPr kinase/phosphorylase with the His6 tag (His-HPrK), produced by E. coli strain BL21(DE3) bearing plasmid pET-hprK, was purified by affinity column chromatography (Ni-nitrilotriacetic acid [NTA]-agarose; Qiagen).

Staining:

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: The HPr protein, produced by E. coli strain BL21(DE3) (Merck Millipore) bearing plasmid pET-ptsH, was purified by one column chromatography step (Toyopearl DEAE-650M). .. The purity of each of the recombinant proteins was evaluated by SDS-PAGE with Coomassie brilliant blue (CBB) staining.

SDS Page:

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: The HPr protein, produced by E. coli strain BL21(DE3) (Merck Millipore) bearing plasmid pET-ptsH, was purified by one column chromatography step (Toyopearl DEAE-650M). .. The purity of each of the recombinant proteins was evaluated by SDS-PAGE with Coomassie brilliant blue (CBB) staining.

Plasmid Preparation:

Article Title: Truncated Hantavirus Nucleocapsid Proteins for Serotyping Sin Nombre, Andes, and Laguna Negra Hantavirus Infections in Humans and Rodents ▿
Article Snippet: .. The DNA fractions were subcloned into the pET43b vector (Merck KGaA, Darmstadt, Germany) and transfected into E. coli strain BL21(DE3) (Merck KGaA). .. A single colony was inoculated into Circle growth medium (MP Biomedicals, Morgan Irvine, CA) containing ampicillin (50 μg/ml) for small-scale culture incubation at 37°C overnight.

Article Title: Efficient Single-Strand Break Repair Requires Binding to Both Poly(ADP-Ribose) and DNA by the Central BRCT Domain of XRCC1
Article Snippet: .. E. coli strain BL21(DE3) (Merck Millipore) was co-transformed with pET15b-SUMO-XRCC1-BRCT1 plasmid. ..

Article Title: Solubilization and renaturation of biologically active human bone morphogenetic protein-4 from inclusion bodies
Article Snippet: .. The expression vector pET24a-hBMP4 was then transformed into the E. coli strain BL21(DE3) (Merck Millipore, USA). ..

Article Title: Histone H1 Recruitment by CHD8 Is Essential for Suppression of the Wnt-?-Catenin Signaling Pathway
Article Snippet: Complementary DNAs encoding wild-type or mutant forms of mouse CHD8S tagged with the FLAG or Myc epitope at the NH2 terminus were subcloned into pcDNA3 (Invitrogen) with the use of the Gateway vector conversion system (Invitrogen); those encoding wild-type or mutant forms of mouse histone H1c with three copies of the FLAG epitope at the NH2 terminus were also subcloned into pcDNA3. .. His6 -tagged proteins were expressed in Escherichia coli strain BL21(DE3)pLys(S) (Merck, Darmstadt, Germany).

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: The other three fragments were digested with XhoI/BamHI and then cloned into the pColdI vector (TaKaRa Bio), which had been treated with the same restriction enzymes, to yield expression plasmids pCold-rhaA, pCold-rhaB, and pCold-EcrhaB. .. E. coli strain BL21 (Merck Millipore), transformed with pCold-rhaR, was grown with shaking in a Luria-Bertani medium ( ) supplemented with ampicillin (50 μg/ml) at 37°C to an OD600 of 0.4.

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: .. The HPr protein, produced by E. coli strain BL21(DE3) (Merck Millipore) bearing plasmid pET-ptsH, was purified by one column chromatography step (Toyopearl DEAE-650M). .. An N-terminal in-frame fusion of B. subtilis HPr kinase/phosphorylase with the His6 tag (His-HPrK), produced by E. coli strain BL21(DE3) bearing plasmid pET-hprK, was purified by affinity column chromatography (Ni-nitrilotriacetic acid [NTA]-agarose; Qiagen).

Article Title: De novo GTP Biosynthesis Is Critical for Virulence of the Fungal Pathogen Cryptococcus neoformans
Article Snippet: Cloning and plasmid propagation was performed in E. coli strain Mach1 (Life Technologies, Carlsbad CA). .. Recombinant Cryptococcus IMPDH was heterologously expressed in E. coli strain BL21(DE3)pLysS (Merck, Darmstadt, Germany).

Article Title: Solubilization and renaturation of biologically active human bone morphogenetic protein-4 from inclusion bodies
Article Snippet: .. The expression vector pET24a-hBMP4 was then transformed into the E. coli strain BL21(DE3) (Merck Millipore, USA). ..

Article Title: The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis
Article Snippet: The amplified product was later inserted into a pET28a vector between Nde I and Xho I restriction sites. .. The recombinant protein with an N-terminal His6 -tag was produced in Escherichia coli strain BL21(DE3) (Merck, Germany) at 22 °C with an incubation for 16–20 h after induction with 0.5 mM isopropyl β-d -1-thiogalactopyranoside (IPTG).

Binding Assay:

Article Title: Truncated Hantavirus Nucleocapsid Proteins for Serotyping Sin Nombre, Andes, and Laguna Negra Hantavirus Infections in Humans and Rodents ▿
Article Snippet: The DNA fractions were subcloned into the pET43b vector (Merck KGaA, Darmstadt, Germany) and transfected into E. coli strain BL21(DE3) (Merck KGaA). .. The cultured cells were collected by centrifugation, resuspended in 5 ml of 0.5 M NaCl binding buffer (0.5 M NaCl, 20 mM imidazole, 20 mM potassium phosphate), and sonicated on ice four times for 15 s each time.

In Vitro:

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: The HPr protein, produced by E. coli strain BL21(DE3) (Merck Millipore) bearing plasmid pET-ptsH, was purified by one column chromatography step (Toyopearl DEAE-650M). .. Ser-46 of the HPr protein was phosphorylated in vitro to form P-Ser-HPr by using the His-HPrK, and then the kinase was removed from the reaction mixture by its passage through an Ni-NTA-agarose column.

Column Chromatography:

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: .. The HPr protein, produced by E. coli strain BL21(DE3) (Merck Millipore) bearing plasmid pET-ptsH, was purified by one column chromatography step (Toyopearl DEAE-650M). .. An N-terminal in-frame fusion of B. subtilis HPr kinase/phosphorylase with the His6 tag (His-HPrK), produced by E. coli strain BL21(DE3) bearing plasmid pET-hprK, was purified by affinity column chromatography (Ni-nitrilotriacetic acid [NTA]-agarose; Qiagen).

Produced:

Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis
Article Snippet: .. The HPr protein, produced by E. coli strain BL21(DE3) (Merck Millipore) bearing plasmid pET-ptsH, was purified by one column chromatography step (Toyopearl DEAE-650M). .. An N-terminal in-frame fusion of B. subtilis HPr kinase/phosphorylase with the His6 tag (His-HPrK), produced by E. coli strain BL21(DE3) bearing plasmid pET-hprK, was purified by affinity column chromatography (Ni-nitrilotriacetic acid [NTA]-agarose; Qiagen).

Article Title: The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis
Article Snippet: .. The recombinant protein with an N-terminal His6 -tag was produced in Escherichia coli strain BL21(DE3) (Merck, Germany) at 22 °C with an incubation for 16–20 h after induction with 0.5 mM isopropyl β-d -1-thiogalactopyranoside (IPTG). .. Harvested cells were resuspended in the lysis buffer containing 50 mM Tris pH 8.0, 500 mM NaCl, 10 mM imidazole, 10 mM β-mercaptoethanol, and 1 mM ZnCl2 .

FLAG-tag:

Article Title: Histone H1 Recruitment by CHD8 Is Essential for Suppression of the Wnt-?-Catenin Signaling Pathway
Article Snippet: Complementary DNAs encoding wild-type or mutant forms of mouse CHD8S tagged with the FLAG or Myc epitope at the NH2 terminus were subcloned into pcDNA3 (Invitrogen) with the use of the Gateway vector conversion system (Invitrogen); those encoding wild-type or mutant forms of mouse histone H1c with three copies of the FLAG epitope at the NH2 terminus were also subcloned into pcDNA3. .. His6 -tagged proteins were expressed in Escherichia coli strain BL21(DE3)pLys(S) (Merck, Darmstadt, Germany).

Lysis:

Article Title: The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis
Article Snippet: The recombinant protein with an N-terminal His6 -tag was produced in Escherichia coli strain BL21(DE3) (Merck, Germany) at 22 °C with an incubation for 16–20 h after induction with 0.5 mM isopropyl β-d -1-thiogalactopyranoside (IPTG). .. Harvested cells were resuspended in the lysis buffer containing 50 mM Tris pH 8.0, 500 mM NaCl, 10 mM imidazole, 10 mM β-mercaptoethanol, and 1 mM ZnCl2 .

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    Merck KGaA bl21 de3 plyss strain
    Small Scale expression analysis from pBDDP-SPR3 constructs. GTPases were expressed in the <t>Bl21</t> (DE3) <t>pLysS</t> strain of E. coli in Luria Bertani broth supplemented with 30 μg/ml kanamycin sulfate. A 1 ml sample of cells was collected before induction and at the time of cell harvesting. This allowed the assessment of total cell protein by SDS-PAGE, and the visualisation of over-expressed target protein. A 40 ml sample of expression culture was applied to a small scale CoNTA IMAC purification to demonstrate the presence of soluble his-tagged protein. (As reported by others, CoNTA spin column purification has proved particularly useful in this respect and gives a consistently clear indication of levels and integrity of recombinant proteins [20] .) In all three cases there was significant soluble expression of the GTPase allowing progression to FPLC scale purification (H 8 -H 8 -KRas 1–169, 24.8 kDa; H 8 -H 8 -Rac1-2-177, 26.2 kDa; H 8 -H 8 -RalB, 25.5 kDa).
    Bl21 De3 Plyss Strain, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 80/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bl21 de3 plyss strain/product/Merck KGaA
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    bl21 de3 plyss strain - by Bioz Stars, 2020-01
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    86
    Merck KGaA e coli bl21 gold de3 strain
    Small Scale expression analysis from pBDDP-SPR3 constructs. GTPases were expressed in the <t>Bl21</t> (DE3) <t>pLysS</t> strain of E. coli in Luria Bertani broth supplemented with 30 μg/ml kanamycin sulfate. A 1 ml sample of cells was collected before induction and at the time of cell harvesting. This allowed the assessment of total cell protein by SDS-PAGE, and the visualisation of over-expressed target protein. A 40 ml sample of expression culture was applied to a small scale CoNTA IMAC purification to demonstrate the presence of soluble his-tagged protein. (As reported by others, CoNTA spin column purification has proved particularly useful in this respect and gives a consistently clear indication of levels and integrity of recombinant proteins [20] .) In all three cases there was significant soluble expression of the GTPase allowing progression to FPLC scale purification (H 8 -H 8 -KRas 1–169, 24.8 kDa; H 8 -H 8 -Rac1-2-177, 26.2 kDa; H 8 -H 8 -RalB, 25.5 kDa).
    E Coli Bl21 Gold De3 Strain, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 86/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Merck KGaA escherichia coli strain bl21
    Small Scale expression analysis from pBDDP-SPR3 constructs. GTPases were expressed in the <t>Bl21</t> (DE3) <t>pLysS</t> strain of E. coli in Luria Bertani broth supplemented with 30 μg/ml kanamycin sulfate. A 1 ml sample of cells was collected before induction and at the time of cell harvesting. This allowed the assessment of total cell protein by SDS-PAGE, and the visualisation of over-expressed target protein. A 40 ml sample of expression culture was applied to a small scale CoNTA IMAC purification to demonstrate the presence of soluble his-tagged protein. (As reported by others, CoNTA spin column purification has proved particularly useful in this respect and gives a consistently clear indication of levels and integrity of recombinant proteins [20] .) In all three cases there was significant soluble expression of the GTPase allowing progression to FPLC scale purification (H 8 -H 8 -KRas 1–169, 24.8 kDa; H 8 -H 8 -Rac1-2-177, 26.2 kDa; H 8 -H 8 -RalB, 25.5 kDa).
    Escherichia Coli Strain Bl21, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Small Scale expression analysis from pBDDP-SPR3 constructs. GTPases were expressed in the Bl21 (DE3) pLysS strain of E. coli in Luria Bertani broth supplemented with 30 μg/ml kanamycin sulfate. A 1 ml sample of cells was collected before induction and at the time of cell harvesting. This allowed the assessment of total cell protein by SDS-PAGE, and the visualisation of over-expressed target protein. A 40 ml sample of expression culture was applied to a small scale CoNTA IMAC purification to demonstrate the presence of soluble his-tagged protein. (As reported by others, CoNTA spin column purification has proved particularly useful in this respect and gives a consistently clear indication of levels and integrity of recombinant proteins [20] .) In all three cases there was significant soluble expression of the GTPase allowing progression to FPLC scale purification (H 8 -H 8 -KRas 1–169, 24.8 kDa; H 8 -H 8 -Rac1-2-177, 26.2 kDa; H 8 -H 8 -RalB, 25.5 kDa).

    Journal: Protein Expression and Purification

    Article Title: A fully automated procedure for the parallel, multidimensional purification and nucleotide loading of the human GTPases KRas, Rac1 and RalB

    doi: 10.1016/j.pep.2017.01.010

    Figure Lengend Snippet: Small Scale expression analysis from pBDDP-SPR3 constructs. GTPases were expressed in the Bl21 (DE3) pLysS strain of E. coli in Luria Bertani broth supplemented with 30 μg/ml kanamycin sulfate. A 1 ml sample of cells was collected before induction and at the time of cell harvesting. This allowed the assessment of total cell protein by SDS-PAGE, and the visualisation of over-expressed target protein. A 40 ml sample of expression culture was applied to a small scale CoNTA IMAC purification to demonstrate the presence of soluble his-tagged protein. (As reported by others, CoNTA spin column purification has proved particularly useful in this respect and gives a consistently clear indication of levels and integrity of recombinant proteins [20] .) In all three cases there was significant soluble expression of the GTPase allowing progression to FPLC scale purification (H 8 -H 8 -KRas 1–169, 24.8 kDa; H 8 -H 8 -Rac1-2-177, 26.2 kDa; H 8 -H 8 -RalB, 25.5 kDa).

    Article Snippet: 2.2 Expression of the CDC25 GEF domain of SOS1 and G-domains of KRas 4B, Rac1 and RalB in pBDDP-SPR3 Plasmids were transformed into the BL21 (DE3) pLysS strain of E. coli (Merck Millipore) and the transformed cells were cultured overnight at 37 °C in Luria Bertani broth supplemented with 30 μg/ml kanamycin sulfate (Melford Laboratories Ltd).

    Techniques: Expressing, Construct, Cell Harvesting, SDS Page, Purification, Recombinant, Fast Protein Liquid Chromatography