e coli bl21 de3 plyss competent cells  (Millipore)


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    Structured Review

    Millipore e coli bl21 de3 plyss competent cells
    SDS-PAGE and WB Analysis of USMTOXO1 expressions in <t>BL21</t> <t>pLysS</t> (DE3), 1 Coomassie blue stained; Lane M : molecular weight marker, Lane 1 : lysate of IPTG-induced E.coli containing vector without insert, Lane 2 : lysate of non-induced E.coli containing vector with insert, Lane 3 : lysate of IPTG-induced E.coli containing vector with insert, Lane 4 : purified USMTOXO1 synthetic protein 2 Western-blot analysis of purified USM.TOXO1; detected by ( A ) anti-His antibody ( B ) human sera
    E Coli Bl21 De3 Plyss Competent Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    e coli bl21 de3 plyss competent cells - by Bioz Stars, 2020-04
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    1) Product Images from "Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans"

    Article Title: Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans

    Journal: Parasites & Vectors

    doi: 10.1186/s13071-015-0932-0

    SDS-PAGE and WB Analysis of USMTOXO1 expressions in BL21 pLysS (DE3), 1 Coomassie blue stained; Lane M : molecular weight marker, Lane 1 : lysate of IPTG-induced E.coli containing vector without insert, Lane 2 : lysate of non-induced E.coli containing vector with insert, Lane 3 : lysate of IPTG-induced E.coli containing vector with insert, Lane 4 : purified USMTOXO1 synthetic protein 2 Western-blot analysis of purified USM.TOXO1; detected by ( A ) anti-His antibody ( B ) human sera
    Figure Legend Snippet: SDS-PAGE and WB Analysis of USMTOXO1 expressions in BL21 pLysS (DE3), 1 Coomassie blue stained; Lane M : molecular weight marker, Lane 1 : lysate of IPTG-induced E.coli containing vector without insert, Lane 2 : lysate of non-induced E.coli containing vector with insert, Lane 3 : lysate of IPTG-induced E.coli containing vector with insert, Lane 4 : purified USMTOXO1 synthetic protein 2 Western-blot analysis of purified USM.TOXO1; detected by ( A ) anti-His antibody ( B ) human sera

    Techniques Used: SDS Page, Western Blot, Staining, Molecular Weight, Marker, Plasmid Preparation, Purification

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    Clone Assay:

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    Centrifugation:

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    Blocking Assay:

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    Incubation:

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    Expressing:

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    Western Blot:

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    Transformation Assay:

    Article Title: The C-terminus of the P22 tailspike protein acts as an independent oligomerization domain for monomeric proteins
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    Article Title: C-terminal hydrophobic interactions play a critical role in oligomeric assembly of the P22 tailspike trimer
    Article Snippet: .. Chemically competent E. coli BL21(DE3) cells (Novagen) were transformed with a pET11a plasmid containing the appropriate gene and selected for on LB-Ampicillin plates. .. Individual colonies were grown in LB media ( ) to an OD600 ~0.5 at 30°C in LB media and induced using 1 mM IPTG for 4 to 24 h. Radioactive protein was produced as described ( ).

    Article Title: Solid-State Nuclear Magnetic Resonance Spectroscopy of Human Immunodeficiency Virus gp41 Protein that Includes the Fusion Peptide: NMR Detection of Recombinant Fgp41 in Inclusion Bodies in Whole Bacterial Cells and Structural Characterization of Purified and Membrane-Associated Fgp41
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    Article Title: Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans
    Article Snippet: .. Consequently, the recombinant plasmid was transformed into E. coli BL21 (DE3) plysS competent cells (Novagen, USA). .. Following the confirmation of the sequences of inserts by DNA sequencing (IDT, Singapore), the protein expression was induced by isopropyl-D thiogalactopyranoside (IPTG) with a final concentration of 1 mM.

    Article Title: Biochemical studies on Francisella tularensis RelA in (p)ppGpp biosynthesis
    Article Snippet: .. Expression of Ft RelA The plasmid pET16b::relA was chemically transformed into E. coli BL21 (DE3) pLysS competent cells (Sigma–Aldrich). .. Single colonies were used to inoculate 2YT media [ ] (10 ml, containing 100 μg/ml ampicillin and 30 μg/ml chloramphenicol) and cultured overnight at 37°C.

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    Article Title: Encapsulation mechanisms and structural studies of GRM2 bacterial microcompartment particles
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    Flow Cytometry:

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    Cell Culture:

    Article Title: Biochemical studies on Francisella tularensis RelA in (p)ppGpp biosynthesis
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    Article Title: Crystallographic Observation of pH-Induced Conformational Changes in the Amyelois transitella Pheromone-Binding Protein AtraPBP1
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    SDS Page:

    Article Title: Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans
    Article Snippet: Consequently, the recombinant plasmid was transformed into E. coli BL21 (DE3) plysS competent cells (Novagen, USA). .. Directly after purification of the synthetic protein by Ni-NTA column, SDS-PAGE and Western blot analysis were carried out to verify the expression of the candidate protein.

    DNA Sequencing:

    Article Title: Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans
    Article Snippet: Consequently, the recombinant plasmid was transformed into E. coli BL21 (DE3) plysS competent cells (Novagen, USA). .. Following the confirmation of the sequences of inserts by DNA sequencing (IDT, Singapore), the protein expression was induced by isopropyl-D thiogalactopyranoside (IPTG) with a final concentration of 1 mM.

    Sequencing:

    Article Title: Solid-State Nuclear Magnetic Resonance Spectroscopy of Human Immunodeficiency Virus gp41 Protein that Includes the Fusion Peptide: NMR Detection of Recombinant Fgp41 in Inclusion Bodies in Whole Bacterial Cells and Structural Characterization of Purified and Membrane-Associated Fgp41
    Article Snippet: The Fgp41 plasmid was constructed within the pET24a(+) vector using the DNA sequence of the Q45D5 primary isolate which is grouped with clade A of HIV-1. .. The plasmid was transformed into BL21(DE3) chemically competent E. coli cells (Novagen, Gibbstown, NJ).

    Article Title: Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans
    Article Snippet: Finally, based on the DNA sequence of the predicted epitopes, a 456 bp synthetic gene (USM.TOXO1) was designed using VNTI computer program software (Life Technologies, USA). .. Consequently, the recombinant plasmid was transformed into E. coli BL21 (DE3) plysS competent cells (Novagen, USA).

    Article Title: Study of a Natural Mutant SHV-Type β-Lactamase, SHV-104, from Klebsiella pneumoniae
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    Sonication:

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    Injection:

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    Recombinant:

    Article Title: A validated antibody panel for the characterization of tau post-translational modifications
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    Article Title: Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans
    Article Snippet: .. Consequently, the recombinant plasmid was transformed into E. coli BL21 (DE3) plysS competent cells (Novagen, USA). .. Following the confirmation of the sequences of inserts by DNA sequencing (IDT, Singapore), the protein expression was induced by isopropyl-D thiogalactopyranoside (IPTG) with a final concentration of 1 mM.

    Article Title: Crystallographic Observation of pH-Induced Conformational Changes in the Amyelois transitella Pheromone-Binding Protein AtraPBP1
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    Article Title: Decoupling the Functional Pleiotropy of Stem Cell Factor by Tuning c-Kit Signaling
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    Molecular Weight:

    Article Title: A PROTEIN SWITCH SENSING SYSTEM FOR THE QUANTIFICATION OF SULFATE
    Article Snippet: The plasmid, pET-28(a)+, and BL21(DE3) chemically competent cells were purchased from Novagen (Madison, WI). .. Gel code blue stain and 3,500 Da molecular weight cut off (MWCO) 3–12 mL Slide-A-Lyzer dialysis cassettes were purchased from Pierce (Rockford, IL).

    DNA Extraction:

    Article Title: A PROTEIN SWITCH SENSING SYSTEM FOR THE QUANTIFICATION OF SULFATE
    Article Snippet: DNA isolation kits, gel extraction kits and Ni-NTA resin were purchased from Qiagen (Valencia, CA). .. The plasmid, pET-28(a)+, and BL21(DE3) chemically competent cells were purchased from Novagen (Madison, WI).

    Ion Exchange Chromatography:

    Article Title: Crystallographic Observation of pH-Induced Conformational Changes in the Amyelois transitella Pheromone-Binding Protein AtraPBP1
    Article Snippet: Crystal preparation and structure determination A pET22-b(+) vector containing DNA encoding mature AtraPBP1 was used to transform BL21 (DE3) competent cells (EMD Chemicals, Novagen, Gibbstown, NJ). .. The recombinant AtraPBP1 was purified by a combination of ion-exchange chromatography and gel-filtration as described previously .

    Isolation:

    Article Title: Improved expression and purification of sigma 1 receptor fused to maltose binding protein by alteration of linker sequence
    Article Snippet: Chemically competent BL21(DE3) and B834 were from Novagen (Merck KGaA, Darmstadt, Germany), BL21(DE3)-RILP was from Stratagene, and C41(DE3) and C43(DE3) were from Lucigen. .. The rare codon supplementation plasmid pRARE2 was isolated from Rosetta 2 cells (Novagen) and then transformed into the appropriate strains [ ].

    Size-exclusion Chromatography:

    Article Title: Decoupling the Functional Pleiotropy of Stem Cell Factor by Tuning c-Kit Signaling
    Article Snippet: The purified proteins were biotinylated in vitro with BirA ligase (produced in house) and then re-purified from the reaction mixture by size exclusion chromatography. .. The SCF constructs were transformed into competent BL21(DE3) E. coli (Novagen).

    Labeling:

    Article Title: Solid-State Nuclear Magnetic Resonance Spectroscopy of Human Immunodeficiency Virus gp41 Protein that Includes the Fusion Peptide: NMR Detection of Recombinant Fgp41 in Inclusion Bodies in Whole Bacterial Cells and Structural Characterization of Purified and Membrane-Associated Fgp41
    Article Snippet: The plasmid was transformed into BL21(DE3) chemically competent E. coli cells (Novagen, Gibbstown, NJ). .. Other reagents and their sources include: Luria-Bertani Broth (LB) medium (Acumedia, Lansing, MI); n -octyl-β-D-thioglucopyranoside (bTOG) and isopropyl-β-D-thiogalactopyranoside (IPTG) (Anatrace, Maumee, OH); 1-palmitoyl-2-oleoyl- sn -glycero-3-phosphocholine (POPC) and 1-palmitoyl-2-oleoyl- sn -glycero-3-[phospho-rac-(1-glycerol)] (sodium salt) (POPG) (Avanti Polar Lipids, Alabaster, AL); isotopically labeled amino acids (Cambridge Isotope Laboratories, Andover, MA).

    Purification:

    Article Title: A validated antibody panel for the characterization of tau post-translational modifications
    Article Snippet: .. Purification of recombinant tau and caspase 3 cleavage The tau 2N4R isoform (aa1–441) and the truncated tau (aa1–421) was recombinantly expressed in E. coli strain BL21(DE3) (Sigma-Aldrich, Cat. No. CMC0014) using pET19b as expression vector. ..

    Article Title: Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans
    Article Snippet: Consequently, the recombinant plasmid was transformed into E. coli BL21 (DE3) plysS competent cells (Novagen, USA). .. Directly after purification of the synthetic protein by Ni-NTA column, SDS-PAGE and Western blot analysis were carried out to verify the expression of the candidate protein.

    Article Title: Study of a Natural Mutant SHV-Type β-Lactamase, SHV-104, from Klebsiella pneumoniae
    Article Snippet: Paragraph title: 2.8. Expression and Purification of SHV-104 ... The corresponding vector, pET26b/SHV-104, was used to transform E. coli BL21 (DE3) (pLysS) competent cells (Novagen Inc., Madison, Wis, USA).

    Article Title: Crystallographic Observation of pH-Induced Conformational Changes in the Amyelois transitella Pheromone-Binding Protein AtraPBP1
    Article Snippet: Crystal preparation and structure determination A pET22-b(+) vector containing DNA encoding mature AtraPBP1 was used to transform BL21 (DE3) competent cells (EMD Chemicals, Novagen, Gibbstown, NJ). .. The recombinant AtraPBP1 was purified by a combination of ion-exchange chromatography and gel-filtration as described previously .

    Article Title: Decoupling the Functional Pleiotropy of Stem Cell Factor by Tuning c-Kit Signaling
    Article Snippet: Paragraph title: Protein expression and purification ... The SCF constructs were transformed into competent BL21(DE3) E. coli (Novagen).

    Article Title: Immune- tolerant elastin-like polypeptides (iTEPs) and their application as CTL vaccine carriers
    Article Snippet: Paragraph title: Production and purification of iTEPs and iTEP fusions ... Competent BL21(DE3) E. coli cells (EMD Chemicals, Inc. USA) were transformed with the pET25b(+) expression vector bearing iTEP or iTEP fusion genes.

    Article Title: Encapsulation mechanisms and structural studies of GRM2 bacterial microcompartment particles
    Article Snippet: .. Protein expression and purification pET-Duet1 plasmids containing cmcABC + cmcD and, optionally, pRSF-Duet1 plasmids containing CutC/CutO/CutF/cmcE were transformed into Escherichia coli BL21-DE3 chemically competent cells (Sigma-Aldrich, cat. No CMC0014). .. Cells were grown in 2xTY medium containing 50 μg/ml ampicillin and (if pRSF-Duet1 vector was used) 30 μg/ml kanamycin.

    Polymerase Chain Reaction:

    Article Title: Study of a Natural Mutant SHV-Type β-Lactamase, SHV-104, from Klebsiella pneumoniae
    Article Snippet: The nucleotides sequences of the PCR-generated fragments were firstly verified by sequencing and secondly cloned into the pET26b (+) vector (Novagen Inc., Madison, Wis, USA). .. The corresponding vector, pET26b/SHV-104, was used to transform E. coli BL21 (DE3) (pLysS) competent cells (Novagen Inc., Madison, Wis, USA).

    Construct:

    Article Title: Solid-State Nuclear Magnetic Resonance Spectroscopy of Human Immunodeficiency Virus gp41 Protein that Includes the Fusion Peptide: NMR Detection of Recombinant Fgp41 in Inclusion Bodies in Whole Bacterial Cells and Structural Characterization of Purified and Membrane-Associated Fgp41
    Article Snippet: The Fgp41 plasmid was constructed within the pET24a(+) vector using the DNA sequence of the Q45D5 primary isolate which is grouped with clade A of HIV-1. .. The plasmid was transformed into BL21(DE3) chemically competent E. coli cells (Novagen, Gibbstown, NJ).

    Article Title: Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans
    Article Snippet: Subsequently, 19 overlapping, single-stranded DNA oligonucleotides (24–44 nt. in length) were designed and used to construct the USM.TOXO1 gene synthetically by assembly PCR, as described by Stemmer [ ]. .. Consequently, the recombinant plasmid was transformed into E. coli BL21 (DE3) plysS competent cells (Novagen, USA).

    Article Title: Decoupling the Functional Pleiotropy of Stem Cell Factor by Tuning c-Kit Signaling
    Article Snippet: .. The SCF constructs were transformed into competent BL21(DE3) E. coli (Novagen). .. Bacterial culture was induced with 1 mM Isopropyl β-D-1-thiogalactopyranoside (IPTG) at 37 °C for 4 h. Inclusion bodies were purified and dissolved in 8 M urea, 50 mM sodium acetate, pH 6.0, 0.1 mM ethylenediamine tetraacetic acid (EDTA, pH 8.0) and 1 mM dithiothreitol (DTT).

    Bradford Protein Assay:

    Article Title: A PROTEIN SWITCH SENSING SYSTEM FOR THE QUANTIFICATION OF SULFATE
    Article Snippet: The Bradford protein assay kit was purchased from Bio-Rad Laboratories, Inc. (Hercules, CA). .. The plasmid, pET-28(a)+, and BL21(DE3) chemically competent cells were purchased from Novagen (Madison, WI).

    Gel Extraction:

    Article Title: A PROTEIN SWITCH SENSING SYSTEM FOR THE QUANTIFICATION OF SULFATE
    Article Snippet: DNA isolation kits, gel extraction kits and Ni-NTA resin were purchased from Qiagen (Valencia, CA). .. The plasmid, pET-28(a)+, and BL21(DE3) chemically competent cells were purchased from Novagen (Madison, WI).

    Polymerase Cycling Assembly:

    Article Title: Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans
    Article Snippet: Subsequently, 19 overlapping, single-stranded DNA oligonucleotides (24–44 nt. in length) were designed and used to construct the USM.TOXO1 gene synthetically by assembly PCR, as described by Stemmer [ ]. .. Consequently, the recombinant plasmid was transformed into E. coli BL21 (DE3) plysS competent cells (Novagen, USA).

    Chromatin Immunoprecipitation:

    Article Title: Decoupling the Functional Pleiotropy of Stem Cell Factor by Tuning c-Kit Signaling
    Article Snippet: To produce recombinant proteins for yeast display selections and immobilization on streptavidin BIAcore chip (see below), mKitD1-3 and hKitD1-3 were expressed with a carboxy-terminal bio acceptor peptide tag (GLNDIFEAQKIEWHE) and purified as described above. .. The SCF constructs were transformed into competent BL21(DE3) E. coli (Novagen).

    Liquid Chromatography:

    Article Title: A validated antibody panel for the characterization of tau post-translational modifications
    Article Snippet: Purification of recombinant tau and caspase 3 cleavage The tau 2N4R isoform (aa1–441) and the truncated tau (aa1–421) was recombinantly expressed in E. coli strain BL21(DE3) (Sigma-Aldrich, Cat. No. CMC0014) using pET19b as expression vector. .. After centrifugation (12 000×g, 4 °C, 15 min) the filtrated supernatant was purified with liquid chromatography using a HiTrap™ IEX HP column (Fisher Scientific, Cat. No. 11748508): The column was washed with sodium-phosphate buffer (50 mM NaPi, 1 mM EGTA, 1 mM DTT, pH 7) and the protein was eluted using a gradient of 0–300 mM NaCl in sodium-phosphate buffer.

    Plasmid Preparation:

    Article Title: Determining Structures of RNA Aptamers and Riboswitches by X-Ray Crystallography
    Article Snippet: .. Chemically competent BL21(DE3) E. coli cells (Novagen, Madison, WI). pHMM expression vector ( see ). ..

    Article Title: A validated antibody panel for the characterization of tau post-translational modifications
    Article Snippet: .. Purification of recombinant tau and caspase 3 cleavage The tau 2N4R isoform (aa1–441) and the truncated tau (aa1–421) was recombinantly expressed in E. coli strain BL21(DE3) (Sigma-Aldrich, Cat. No. CMC0014) using pET19b as expression vector. ..

    Article Title: The C-terminus of the P22 tailspike protein acts as an independent oligomerization domain for monomeric proteins
    Article Snippet: .. Chemically competent E. coli BL21(DE3) cells (Novagen) were transformed with the pMal-c2g plasmid containing the appropriate gene and selected for on LB + Amp plates. ..

    Article Title: C-terminal hydrophobic interactions play a critical role in oligomeric assembly of the P22 tailspike trimer
    Article Snippet: .. Chemically competent E. coli BL21(DE3) cells (Novagen) were transformed with a pET11a plasmid containing the appropriate gene and selected for on LB-Ampicillin plates. .. Individual colonies were grown in LB media ( ) to an OD600 ~0.5 at 30°C in LB media and induced using 1 mM IPTG for 4 to 24 h. Radioactive protein was produced as described ( ).

    Article Title: Magnetization of active inclusion bodies: comparison with centrifugation in repetitive biotransformations
    Article Snippet: Chemically competent Escherichia coli BL21(DE3)T1R (CMC0014), CelLytic™ B Cell Lysis Reagent, ferrous sulphate heptahydrate, KOH and other compounds were supplied by Sigma-Aldrich (St. Louis, Missouri, USA). .. GFP (FPbase: TurboGFP; GenBank: ), sialic acid aldolase (SAA, UniProt: ; NCBI-GeneID: 947742) and UDP–glucose pyrophosphorylase (GalU, UniProt: ; NCBI-GeneID: 945730) genes were N -terminally fused with the cellulose-binding domain from Clostridium cellulovorans by cloning into plasmid pET-34b (Additional file ).

    Article Title: Solid-State Nuclear Magnetic Resonance Spectroscopy of Human Immunodeficiency Virus gp41 Protein that Includes the Fusion Peptide: NMR Detection of Recombinant Fgp41 in Inclusion Bodies in Whole Bacterial Cells and Structural Characterization of Purified and Membrane-Associated Fgp41
    Article Snippet: .. The plasmid was transformed into BL21(DE3) chemically competent E. coli cells (Novagen, Gibbstown, NJ). .. The Fgp41 amino acid sequence is given in and the correlative sequence from plasmid DNA extracted from the cells is provided in the .

    Article Title: Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans
    Article Snippet: .. Consequently, the recombinant plasmid was transformed into E. coli BL21 (DE3) plysS competent cells (Novagen, USA). .. Following the confirmation of the sequences of inserts by DNA sequencing (IDT, Singapore), the protein expression was induced by isopropyl-D thiogalactopyranoside (IPTG) with a final concentration of 1 mM.

    Article Title: A PROTEIN SWITCH SENSING SYSTEM FOR THE QUANTIFICATION OF SULFATE
    Article Snippet: .. The plasmid, pET-28(a)+, and BL21(DE3) chemically competent cells were purchased from Novagen (Madison, WI). .. Gel code blue stain and 3,500 Da molecular weight cut off (MWCO) 3–12 mL Slide-A-Lyzer dialysis cassettes were purchased from Pierce (Rockford, IL).

    Article Title: Biochemical studies on Francisella tularensis RelA in (p)ppGpp biosynthesis
    Article Snippet: .. Expression of Ft RelA The plasmid pET16b::relA was chemically transformed into E. coli BL21 (DE3) pLysS competent cells (Sigma–Aldrich). .. Single colonies were used to inoculate 2YT media [ ] (10 ml, containing 100 μg/ml ampicillin and 30 μg/ml chloramphenicol) and cultured overnight at 37°C.

    Article Title: Study of a Natural Mutant SHV-Type β-Lactamase, SHV-104, from Klebsiella pneumoniae
    Article Snippet: .. The corresponding vector, pET26b/SHV-104, was used to transform E. coli BL21 (DE3) (pLysS) competent cells (Novagen Inc., Madison, Wis, USA). .. The SHV-104 enzyme was produced in Terrific Broth (TB) medium containing kanamycin (50 μ g/mL) and chloramphenicol (30 μ g/mL) as selecting agents during growth of the bacteria at 37°C under orbital shaking.

    Article Title: Crystallographic Observation of pH-Induced Conformational Changes in the Amyelois transitella Pheromone-Binding Protein AtraPBP1
    Article Snippet: .. Crystal preparation and structure determination A pET22-b(+) vector containing DNA encoding mature AtraPBP1 was used to transform BL21 (DE3) competent cells (EMD Chemicals, Novagen, Gibbstown, NJ). .. The transformant was used to inoculate LB medium containing carbenicilin and cells were cultured at 200 rpm at 28°C overnight.

    Article Title: Immune- tolerant elastin-like polypeptides (iTEPs) and their application as CTL vaccine carriers
    Article Snippet: .. Competent BL21(DE3) E. coli cells (EMD Chemicals, Inc. USA) were transformed with the pET25b(+) expression vector bearing iTEP or iTEP fusion genes. ..

    Article Title: Encapsulation mechanisms and structural studies of GRM2 bacterial microcompartment particles
    Article Snippet: Protein expression and purification pET-Duet1 plasmids containing cmcABC + cmcD and, optionally, pRSF-Duet1 plasmids containing CutC/CutO/CutF/cmcE were transformed into Escherichia coli BL21-DE3 chemically competent cells (Sigma-Aldrich, cat. No CMC0014). .. Cells were grown in 2xTY medium containing 50 μg/ml ampicillin and (if pRSF-Duet1 vector was used) 30 μg/ml kanamycin.

    Software:

    Article Title: Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans
    Article Snippet: Finally, based on the DNA sequence of the predicted epitopes, a 456 bp synthetic gene (USM.TOXO1) was designed using VNTI computer program software (Life Technologies, USA). .. Consequently, the recombinant plasmid was transformed into E. coli BL21 (DE3) plysS competent cells (Novagen, USA).

    Positron Emission Tomography:

    Article Title: Magnetization of active inclusion bodies: comparison with centrifugation in repetitive biotransformations
    Article Snippet: Chemically competent Escherichia coli BL21(DE3)T1R (CMC0014), CelLytic™ B Cell Lysis Reagent, ferrous sulphate heptahydrate, KOH and other compounds were supplied by Sigma-Aldrich (St. Louis, Missouri, USA). .. GFP (FPbase: TurboGFP; GenBank: ), sialic acid aldolase (SAA, UniProt: ; NCBI-GeneID: 947742) and UDP–glucose pyrophosphorylase (GalU, UniProt: ; NCBI-GeneID: 945730) genes were N -terminally fused with the cellulose-binding domain from Clostridium cellulovorans by cloning into plasmid pET-34b (Additional file ).

    Article Title: Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans
    Article Snippet: The synthetic gene was then cloned into a pET-32a expression vector (Novagen, USA). .. Consequently, the recombinant plasmid was transformed into E. coli BL21 (DE3) plysS competent cells (Novagen, USA).

    Article Title: A PROTEIN SWITCH SENSING SYSTEM FOR THE QUANTIFICATION OF SULFATE
    Article Snippet: .. The plasmid, pET-28(a)+, and BL21(DE3) chemically competent cells were purchased from Novagen (Madison, WI). .. Gel code blue stain and 3,500 Da molecular weight cut off (MWCO) 3–12 mL Slide-A-Lyzer dialysis cassettes were purchased from Pierce (Rockford, IL).

    Article Title: Encapsulation mechanisms and structural studies of GRM2 bacterial microcompartment particles
    Article Snippet: .. Protein expression and purification pET-Duet1 plasmids containing cmcABC + cmcD and, optionally, pRSF-Duet1 plasmids containing CutC/CutO/CutF/cmcE were transformed into Escherichia coli BL21-DE3 chemically competent cells (Sigma-Aldrich, cat. No CMC0014). .. Cells were grown in 2xTY medium containing 50 μg/ml ampicillin and (if pRSF-Duet1 vector was used) 30 μg/ml kanamycin.

    In Vitro:

    Article Title: Decoupling the Functional Pleiotropy of Stem Cell Factor by Tuning c-Kit Signaling
    Article Snippet: The purified proteins were biotinylated in vitro with BirA ligase (produced in house) and then re-purified from the reaction mixture by size exclusion chromatography. .. The SCF constructs were transformed into competent BL21(DE3) E. coli (Novagen).

    Produced:

    Article Title: C-terminal hydrophobic interactions play a critical role in oligomeric assembly of the P22 tailspike trimer
    Article Snippet: Chemically competent E. coli BL21(DE3) cells (Novagen) were transformed with a pET11a plasmid containing the appropriate gene and selected for on LB-Ampicillin plates. .. Individual colonies were grown in LB media ( ) to an OD600 ~0.5 at 30°C in LB media and induced using 1 mM IPTG for 4 to 24 h. Radioactive protein was produced as described ( ).

    Article Title: Study of a Natural Mutant SHV-Type β-Lactamase, SHV-104, from Klebsiella pneumoniae
    Article Snippet: The corresponding vector, pET26b/SHV-104, was used to transform E. coli BL21 (DE3) (pLysS) competent cells (Novagen Inc., Madison, Wis, USA). .. The SHV-104 enzyme was produced in Terrific Broth (TB) medium containing kanamycin (50 μ g/mL) and chloramphenicol (30 μ g/mL) as selecting agents during growth of the bacteria at 37°C under orbital shaking.

    Article Title: Decoupling the Functional Pleiotropy of Stem Cell Factor by Tuning c-Kit Signaling
    Article Snippet: The purified proteins were biotinylated in vitro with BirA ligase (produced in house) and then re-purified from the reaction mixture by size exclusion chromatography. .. The SCF constructs were transformed into competent BL21(DE3) E. coli (Novagen).

    Concentration Assay:

    Article Title: Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans
    Article Snippet: Consequently, the recombinant plasmid was transformed into E. coli BL21 (DE3) plysS competent cells (Novagen, USA). .. Following the confirmation of the sequences of inserts by DNA sequencing (IDT, Singapore), the protein expression was induced by isopropyl-D thiogalactopyranoside (IPTG) with a final concentration of 1 mM.

    Article Title: Biochemical studies on Francisella tularensis RelA in (p)ppGpp biosynthesis
    Article Snippet: Expression of Ft RelA The plasmid pET16b::relA was chemically transformed into E. coli BL21 (DE3) pLysS competent cells (Sigma–Aldrich). .. The overnight culture was used as a 1% inoculum for flasks of 2YT (4×1.25 litre) which were induced with IPTG (final concentration of 0.4 mM) when the absorbance at 600 nm (A 600 ) reached 0.6 and then cultured overnight at 16°C.

    Article Title: Study of a Natural Mutant SHV-Type β-Lactamase, SHV-104, from Klebsiella pneumoniae
    Article Snippet: The corresponding vector, pET26b/SHV-104, was used to transform E. coli BL21 (DE3) (pLysS) competent cells (Novagen Inc., Madison, Wis, USA). .. At an absorbance value of 0.7 at 600 nm, IPTG (final concentration 0.5 mM) was added and the culture was incubated for 5 additional hours.

    Lysis:

    Article Title: The C-terminus of the P22 tailspike protein acts as an independent oligomerization domain for monomeric proteins
    Article Snippet: Chemically competent E. coli BL21(DE3) cells (Novagen) were transformed with the pMal-c2g plasmid containing the appropriate gene and selected for on LB + Amp plates. .. The overnight starter culture was used to inoculate a 1.5 litre culture that was grown to a D 600 ~ 0.6 at 37 °C and induced using 1 mM IPTG (isopropyl β- d -thiogalactoside) at 30 °C for 24 h. Cells were harvested by centrifugation (4200 g for 30 min at 4 °C) and resuspended in lysis buffer (50 mM Tris, 5 mM MgCl2 , 0.1 % Triton X-100, 0.1 mg/ml lysozyme and 0.1 mg/ml DNase, pH 7.4).

    Article Title: C-terminal hydrophobic interactions play a critical role in oligomeric assembly of the P22 tailspike trimer
    Article Snippet: Chemically competent E. coli BL21(DE3) cells (Novagen) were transformed with a pET11a plasmid containing the appropriate gene and selected for on LB-Ampicillin plates. .. Cells were harvested by centrifugation and resuspended in lysis buffer (50 mM Tris, 5 mM MgCl2 , 0.1% Triton X-100, 0.1 mg/mL lysozyme, 0.1 mg/mL DNase).

    Article Title: Magnetization of active inclusion bodies: comparison with centrifugation in repetitive biotransformations
    Article Snippet: .. Chemically competent Escherichia coli BL21(DE3)T1R (CMC0014), CelLytic™ B Cell Lysis Reagent, ferrous sulphate heptahydrate, KOH and other compounds were supplied by Sigma-Aldrich (St. Louis, Missouri, USA). .. GFP (FPbase: TurboGFP; GenBank: ), sialic acid aldolase (SAA, UniProt: ; NCBI-GeneID: 947742) and UDP–glucose pyrophosphorylase (GalU, UniProt: ; NCBI-GeneID: 945730) genes were N -terminally fused with the cellulose-binding domain from Clostridium cellulovorans by cloning into plasmid pET-34b (Additional file ).

    Article Title: Encapsulation mechanisms and structural studies of GRM2 bacterial microcompartment particles
    Article Snippet: Protein expression and purification pET-Duet1 plasmids containing cmcABC + cmcD and, optionally, pRSF-Duet1 plasmids containing CutC/CutO/CutF/cmcE were transformed into Escherichia coli BL21-DE3 chemically competent cells (Sigma-Aldrich, cat. No CMC0014). .. Cell lysis was performed in a buffer containing 100 mM Tris-HCl (pH 8.0), 300 mM NaCl, 0.1% Triton X-100, 20 mm MgSO4 , 0.1 mg/ml DNase, 1 mg/ml lysozyme, 1 mM PMSF, and 2 mM DTT.

    Staining:

    Article Title: A PROTEIN SWITCH SENSING SYSTEM FOR THE QUANTIFICATION OF SULFATE
    Article Snippet: The plasmid, pET-28(a)+, and BL21(DE3) chemically competent cells were purchased from Novagen (Madison, WI). .. Gel code blue stain and 3,500 Da molecular weight cut off (MWCO) 3–12 mL Slide-A-Lyzer dialysis cassettes were purchased from Pierce (Rockford, IL).

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    Millipore e coli bl21 de3 plyss prsetc cells
    E Coli Bl21 De3 Plyss Prsetc Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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