bl21 star de3 competent cells  (Thermo Fisher)


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    Structured Review

    Thermo Fisher bl21 star de3 competent cells
    Bl21 Star De3 Competent Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bl21 star de3 competent cells/product/Thermo Fisher
    Average 99 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    bl21 star de3 competent cells - by Bioz Stars, 2020-02
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Plasticity, dynamics, and inhibition of emerging tetracycline-resistance enzymes
    Article Snippet: Paragraph title: Cloning, expression and purification of tetracycline-inactivating enzymes ... Constructs were transformed into BL21-Star (DE3) competent cells (Life Technologies).

    Article Title: A dual purpose universal influenza vaccine candidate confers protective immunity against anthrax
    Article Snippet: For EFn‐M2, the EFn sequence was cloned upstream of the full‐length M2 sequence in the pET200/D‐TOPO plasmid. .. All expression plasmids were transformed in BL21 Star™ (DE3) competent cells (Life Technologies).

    Article Title: Chemerin158K Protein Is the Dominant Chemerin Isoform in Synovial and Cerebrospinal Fluids but Not in Plasma *
    Article Snippet: .. cDNA (Open Biosystems, Huntsville, AL) encoding human chem163S, chem158K, and chem157S lacking the 20-amino acid signal peptide were cloned into pGEX 6P-3 vector (GE Healthcare) and transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The resultant expression plasmids were verified by sequencing.

    Article Title: De novo design of a homo-trimeric amantadine-binding protein
    Article Snippet: Paragraph title: Cloning, protein expression and purification ... Constructs were transformed into BL21-Star (DE3) competent cells (Life Technologies).

    Centrifugation:

    Article Title: Structural Insights into c-Myc-interacting Zinc Finger Protein-1 (Miz-1) Delineate Domains Required for DNA Scanning and Sequence-specific Binding *
    Article Snippet: The construct was transformed in BL21 star (DE3) competent cells (Invitrogen). .. Bacteria were grown in M9 medium containing 15 NH4 Cl and [13 C]glucose to an A 600 nm of 0.6, were induced for 15 h at 37 °C with 0.5 m m isopropyl 1-thio-β- d -galactopyranoside, and were harvested by centrifugation.

    Amplification:

    Article Title: Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM+ B Cells in The Absence of Germinal Centers
    Article Snippet: Production of rainbow trout recombinant IL-6 The sequence encoding the mature peptide of trout IL-6 was amplified from spleen cDNA prepared from Aeromonas salmonicida infected fish and cloned to the pET/Duet-1 vector (Novagen, UK). .. A plasmid was used to transform BL21 Star (DE3) competent cells (Invitrogen) and protein expression was induced by IPTG and purified under denaturing conditions as described previously .

    Synthesized:

    Article Title: A dual purpose universal influenza vaccine candidate confers protective immunity against anthrax
    Article Snippet: The full‐length, avian influenza A (H5N1) M2 DNA sequence was synthesized by Retrogen, Inc. (San Diego, CA). .. All expression plasmids were transformed in BL21 Star™ (DE3) competent cells (Life Technologies).

    Construct:

    Article Title: Evolution of Th2 responses: characterization of IL-4/13 in sea bass (Dicentrarchus labrax L.) and studies of expression and biological activity
    Article Snippet: The constructs encode identical mature peptides with a His-tag (MGSHHHHHHHHS) added at the N-terminus for easy purification. .. Following transformation of the plasmid into BL21 Star (DE3) competent cells (Invitrogen) and the induction of recombinant protein production, a first purification was performed under denaturing conditions.

    Article Title: Plasticity, dynamics, and inhibition of emerging tetracycline-resistance enzymes
    Article Snippet: .. Constructs were transformed into BL21-Star (DE3) competent cells (Life Technologies). ..

    Article Title: Structural Insights into c-Myc-interacting Zinc Finger Protein-1 (Miz-1) Delineate Domains Required for DNA Scanning and Sequence-specific Binding *
    Article Snippet: .. The construct was transformed in BL21 star (DE3) competent cells (Invitrogen). .. Bacteria were grown in M9 medium containing 15 NH4 Cl and [13 C]glucose to an A 600 nm of 0.6, were induced for 15 h at 37 °C with 0.5 m m isopropyl 1-thio-β- d -galactopyranoside, and were harvested by centrifugation.

    Article Title: A mutagenesis screen for essential plastid biogenesis genes in human malaria parasites
    Article Snippet: BL21 Star (DE3) competent cells (Thermo Fisher) were used for the WT condition. .. For each construct, a colony was picked and washed in M9 minimal media (M9) (22 mM potassium phosphate monobasic, 22 mM sodium phosphate dibasic, 85 mM sodium chloride, 18.7 mM ammonium chloride, 2 mM magnesium sulfate, 0.1 mM calcium chloride, and 0.4% glycerol), resuspended in M9, and streaked onto M9/agar plates containing either carbenicillin (100 μg/mL) or carbenicillin and 1 mM L-tryptophan (Sigma).

    Article Title: Studies on the Use of Flagellin as an Immunostimulant and Vaccine Adjuvant in Fish Aquaculture
    Article Snippet: The mutant YRF constructs, YRF-N, YRF-C and YRF-NC, were prepared from pTRI-YRF by PCR using the Q5 high fidelity enzyme (New England Biolabs, United Kingdom) and re-ligation, using primer pairs GCCAGTTCCGCTCATCACCAC/GGAACGGAAGTTACCGTTAACCATC (YRF-N), GCCCATGGTATATCTCCTTTGATTGT/GATAACCGCACGGCAGCCA (YRF-C), and CAAGACTTTAATGCCGTTGAAATCGGT/GTTGAAGCCAAAGGTTTTGACGTATTGA (YRF-NC), respectively. .. Following sequence confirmation, the transformation of BL21 Star (DE3) competent cells (Invitrogen), induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously ( ).

    Article Title: Rainbow trout CK9, a CCL25-like ancient chemokine that attracts and regulates B cells and macrophages, the main antigen presenting cells in fish
    Article Snippet: The construct (pET-CK9) encodes an identical CK9 mature amino acid sequence with the addition of methionine at the N-terminus and a his-tag (GSGHHHHHHHH) added at the C-terminus for purification. .. Following transformation of the pET-CK9 plasmid into BL21 Star (DE3) competent cells (Invitrogen), the induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ].

    Article Title: De novo design of a homo-trimeric amantadine-binding protein
    Article Snippet: .. Constructs were transformed into BL21-Star (DE3) competent cells (Life Technologies). .. Cells harboring the plasmid were grown at 37°C in Terrific Broth II medium containing a final concentration of 0.05 mg/ml kanamycin.

    Cell Culture:

    Article Title: A dual purpose universal influenza vaccine candidate confers protective immunity against anthrax
    Article Snippet: All expression plasmids were transformed in BL21 Star™ (DE3) competent cells (Life Technologies). .. Transformed cells were cultured in Luria–Bertani medium with 50 μg/ml of kanamycin to an optical density at 600 nm of 0·5, and expression was induced with 0·001–1 m m Isopropyl β ‐ d ‐1‐thiogalactopyranoside depending on the protein, and harvested 2–4 hr later.

    Expressing:

    Article Title: Plasticity, dynamics, and inhibition of emerging tetracycline-resistance enzymes
    Article Snippet: Paragraph title: Cloning, expression and purification of tetracycline-inactivating enzymes ... Constructs were transformed into BL21-Star (DE3) competent cells (Life Technologies).

    Article Title: Arylfluorosulfates Inactivate Intracellular Lipid Binding Protein(s) through Chemoselective SuFEx Reaction with a Binding-site Tyr Residue
    Article Snippet: Paragraph title: Protein expression and purification ... The pET22b vectors encoding FABP5, FABP4, FABP3 and CRABP2 variants were transformed into BL21 Star (DE3) competent cells (Invitrogen).

    Article Title: A dual purpose universal influenza vaccine candidate confers protective immunity against anthrax
    Article Snippet: .. All expression plasmids were transformed in BL21 Star™ (DE3) competent cells (Life Technologies). .. Transformed cells were cultured in Luria–Bertani medium with 50 μg/ml of kanamycin to an optical density at 600 nm of 0·5, and expression was induced with 0·001–1 m m Isopropyl β ‐ d ‐1‐thiogalactopyranoside depending on the protein, and harvested 2–4 hr later.

    Article Title: First in-depth analysis of the novel Th2-type cytokines in salmonid fish reveals distinct patterns of expression and modulation but overlapping bioactivities
    Article Snippet: .. Expression and purification of trout IL-4/13 isoforms For each protein a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ].

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: Paragraph title: Expression and Purification of Trout IL-2 Isoforms ... For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen).

    Article Title: Chemerin158K Protein Is the Dominant Chemerin Isoform in Synovial and Cerebrospinal Fluids but Not in Plasma *
    Article Snippet: cDNA (Open Biosystems, Huntsville, AL) encoding human chem163S, chem158K, and chem157S lacking the 20-amino acid signal peptide were cloned into pGEX 6P-3 vector (GE Healthcare) and transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The resultant expression plasmids were verified by sequencing.

    Article Title: Studies on the Use of Flagellin as an Immunostimulant and Vaccine Adjuvant in Fish Aquaculture
    Article Snippet: The construct pTri-YRF for expression of full-length recombinant Y. ruckeri flagellin (YRF) and the production of YRF was described previously ( ). .. Following sequence confirmation, the transformation of BL21 Star (DE3) competent cells (Invitrogen), induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously ( ).

    Article Title: Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM+ B Cells in The Absence of Germinal Centers
    Article Snippet: .. A plasmid was used to transform BL21 Star (DE3) competent cells (Invitrogen) and protein expression was induced by IPTG and purified under denaturing conditions as described previously . .. The purified, denatured rtIL-6 was refolded in refolding buffer containing 50 mM Tris–HCl, pH8.0, 0.5 M arginine, 0.5% Triton-100 and 5 mM β-mercaptoethanol at 4 °C for 2 days.

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: .. Expression and Purification of Trout IL-2 Isoforms For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins, and quantification of protein concentration were as described previously ( , , ).

    Article Title: De novo design of a homo-trimeric amantadine-binding protein
    Article Snippet: Paragraph title: Cloning, protein expression and purification ... Constructs were transformed into BL21-Star (DE3) competent cells (Life Technologies).

    Transformation Assay:

    Article Title: Evolution of Th2 responses: characterization of IL-4/13 in sea bass (Dicentrarchus labrax L.) and studies of expression and biological activity
    Article Snippet: .. Following transformation of the plasmid into BL21 Star (DE3) competent cells (Invitrogen) and the induction of recombinant protein production, a first purification was performed under denaturing conditions. .. Successively we undertook a refolding process, a re-purification under native conditions, a SDS-PAGE analysis of the purified proteins and a quantification of the protein concentration as described in detail previously – .

    Article Title: Plasticity, dynamics, and inhibition of emerging tetracycline-resistance enzymes
    Article Snippet: .. Constructs were transformed into BL21-Star (DE3) competent cells (Life Technologies). ..

    Article Title: Arylfluorosulfates Inactivate Intracellular Lipid Binding Protein(s) through Chemoselective SuFEx Reaction with a Binding-site Tyr Residue
    Article Snippet: .. The pET22b vectors encoding FABP5, FABP4, FABP3 and CRABP2 variants were transformed into BL21 Star (DE3) competent cells (Invitrogen). .. When the bacteria reached an OD600 of 0.6–0.8, protein expression was induced with 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG, BioPineer Inc.) for 4 h at 37 °C.

    Article Title: A dual purpose universal influenza vaccine candidate confers protective immunity against anthrax
    Article Snippet: .. All expression plasmids were transformed in BL21 Star™ (DE3) competent cells (Life Technologies). .. Transformed cells were cultured in Luria–Bertani medium with 50 μg/ml of kanamycin to an optical density at 600 nm of 0·5, and expression was induced with 0·001–1 m m Isopropyl β ‐ d ‐1‐thiogalactopyranoside depending on the protein, and harvested 2–4 hr later.

    Article Title: Structural Insights into c-Myc-interacting Zinc Finger Protein-1 (Miz-1) Delineate Domains Required for DNA Scanning and Sequence-specific Binding *
    Article Snippet: .. The construct was transformed in BL21 star (DE3) competent cells (Invitrogen). .. Bacteria were grown in M9 medium containing 15 NH4 Cl and [13 C]glucose to an A 600 nm of 0.6, were induced for 15 h at 37 °C with 0.5 m m isopropyl 1-thio-β- d -galactopyranoside, and were harvested by centrifugation.

    Article Title: First in-depth analysis of the novel Th2-type cytokines in salmonid fish reveals distinct patterns of expression and modulation but overlapping bioactivities
    Article Snippet: .. Expression and purification of trout IL-4/13 isoforms For each protein a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ].

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: .. For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins, and quantification of protein concentration were as described previously ( , , ).

    Article Title: Chemerin158K Protein Is the Dominant Chemerin Isoform in Synovial and Cerebrospinal Fluids but Not in Plasma *
    Article Snippet: .. cDNA (Open Biosystems, Huntsville, AL) encoding human chem163S, chem158K, and chem157S lacking the 20-amino acid signal peptide were cloned into pGEX 6P-3 vector (GE Healthcare) and transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The resultant expression plasmids were verified by sequencing.

    Article Title: A mutagenesis screen for essential plastid biogenesis genes in human malaria parasites
    Article Snippet: BL21 Star (DE3) competent cells (Thermo Fisher) were used for the WT condition. .. The competent W3110trpC9800 cells were transformed with the pGEXT vectors containing the different Vitrella or Plasmodium IGPS and IGPS-like genes and were plated on LB agar plates containing carbenicillin.

    Article Title: Studies on the Use of Flagellin as an Immunostimulant and Vaccine Adjuvant in Fish Aquaculture
    Article Snippet: .. Following sequence confirmation, the transformation of BL21 Star (DE3) competent cells (Invitrogen), induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously ( ). .. The monocyte/macrophage-like cell line, RTS-11, from rainbow trout spleen was cultured in Leibovitz (L-15) medium (Invitrogen, United Kingdom) plus 30% fetal calf serum (FCS; Labtech International, United Kingdom) and antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin; Invitrogen, UK) at 20°C, and passaged as described previously ( ).

    Article Title: Rainbow trout CK9, a CCL25-like ancient chemokine that attracts and regulates B cells and macrophages, the main antigen presenting cells in fish
    Article Snippet: .. Following transformation of the pET-CK9 plasmid into BL21 Star (DE3) competent cells (Invitrogen), the induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ]. .. The refolding buffer contained 50 mM Tris-HCl (pH7.5), 10% glycerol, 0.6 M arginine monohydrochloride, 1 M 3-(1-Pyridinio)-1-propanesulfonate (known as NDSB 201 and PPS, Sigma), 0.2% PEG3350 and 5 mM 2-mercaptoethanol.

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: .. Expression and Purification of Trout IL-2 Isoforms For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins, and quantification of protein concentration were as described previously ( , , ).

    Article Title: De novo design of a homo-trimeric amantadine-binding protein
    Article Snippet: .. Constructs were transformed into BL21-Star (DE3) competent cells (Life Technologies). .. Cells harboring the plasmid were grown at 37°C in Terrific Broth II medium containing a final concentration of 0.05 mg/ml kanamycin.

    Protease Inhibitor:

    Article Title: Chemerin158K Protein Is the Dominant Chemerin Isoform in Synovial and Cerebrospinal Fluids but Not in Plasma *
    Article Snippet: cDNA (Open Biosystems, Huntsville, AL) encoding human chem163S, chem158K, and chem157S lacking the 20-amino acid signal peptide were cloned into pGEX 6P-3 vector (GE Healthcare) and transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The proteins were produced in 2× YT medium at 37 °C for 4 h. Cells were pelleted before suspension in STE buffer (50 m m Tris-HCl, 150 m m NaCl, 1 m m EDTA, pH 7.5) containing 1 m m DTT, 5 units/ml rLysozyme (Novagen, Madison, WI), benzonase nuclease, and Complete protease inhibitor (Roche Applied Science).

    Infection:

    Article Title: Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM+ B Cells in The Absence of Germinal Centers
    Article Snippet: Production of rainbow trout recombinant IL-6 The sequence encoding the mature peptide of trout IL-6 was amplified from spleen cDNA prepared from Aeromonas salmonicida infected fish and cloned to the pET/Duet-1 vector (Novagen, UK). .. A plasmid was used to transform BL21 Star (DE3) competent cells (Invitrogen) and protein expression was induced by IPTG and purified under denaturing conditions as described previously .

    Generated:

    Article Title: Structural Insights into c-Myc-interacting Zinc Finger Protein-1 (Miz-1) Delineate Domains Required for DNA Scanning and Sequence-specific Binding *
    Article Snippet: The cDNA of ZFs 1–4 of Miz-1 (Miz1–4, residues 304–414) was generated by PCR from the complete cDNA of Miz-1 using primers F (5′-CAT ATG GTC ATC CAC AAG TGC GAG GAC TGT GG-3′) and R (5′-GGA TCC CTA GCC GCT GTG CAC CAG CTG GTG-3′). .. The construct was transformed in BL21 star (DE3) competent cells (Invitrogen).

    Protein Concentration:

    Article Title: Evolution of Th2 responses: characterization of IL-4/13 in sea bass (Dicentrarchus labrax L.) and studies of expression and biological activity
    Article Snippet: Following transformation of the plasmid into BL21 Star (DE3) competent cells (Invitrogen) and the induction of recombinant protein production, a first purification was performed under denaturing conditions. .. Successively we undertook a refolding process, a re-purification under native conditions, a SDS-PAGE analysis of the purified proteins and a quantification of the protein concentration as described in detail previously – .

    Article Title: First in-depth analysis of the novel Th2-type cytokines in salmonid fish reveals distinct patterns of expression and modulation but overlapping bioactivities
    Article Snippet: Expression and purification of trout IL-4/13 isoforms For each protein a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ].

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins, and quantification of protein concentration were as described previously ( , , ).

    Article Title: Studies on the Use of Flagellin as an Immunostimulant and Vaccine Adjuvant in Fish Aquaculture
    Article Snippet: .. Following sequence confirmation, the transformation of BL21 Star (DE3) competent cells (Invitrogen), induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously ( ). .. The monocyte/macrophage-like cell line, RTS-11, from rainbow trout spleen was cultured in Leibovitz (L-15) medium (Invitrogen, United Kingdom) plus 30% fetal calf serum (FCS; Labtech International, United Kingdom) and antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin; Invitrogen, UK) at 20°C, and passaged as described previously ( ).

    Article Title: Rainbow trout CK9, a CCL25-like ancient chemokine that attracts and regulates B cells and macrophages, the main antigen presenting cells in fish
    Article Snippet: .. Following transformation of the pET-CK9 plasmid into BL21 Star (DE3) competent cells (Invitrogen), the induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ]. .. The refolding buffer contained 50 mM Tris-HCl (pH7.5), 10% glycerol, 0.6 M arginine monohydrochloride, 1 M 3-(1-Pyridinio)-1-propanesulfonate (known as NDSB 201 and PPS, Sigma), 0.2% PEG3350 and 5 mM 2-mercaptoethanol.

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: Expression and Purification of Trout IL-2 Isoforms For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins, and quantification of protein concentration were as described previously ( , , ).

    Sequencing:

    Article Title: A dual purpose universal influenza vaccine candidate confers protective immunity against anthrax
    Article Snippet: For EFn‐M2, the EFn sequence was cloned upstream of the full‐length M2 sequence in the pET200/D‐TOPO plasmid. .. All expression plasmids were transformed in BL21 Star™ (DE3) competent cells (Life Technologies).

    Article Title: First in-depth analysis of the novel Th2-type cytokines in salmonid fish reveals distinct patterns of expression and modulation but overlapping bioactivities
    Article Snippet: .. Expression and purification of trout IL-4/13 isoforms For each protein a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ].

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: .. For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins, and quantification of protein concentration were as described previously ( , , ).

    Article Title: Chemerin158K Protein Is the Dominant Chemerin Isoform in Synovial and Cerebrospinal Fluids but Not in Plasma *
    Article Snippet: cDNA (Open Biosystems, Huntsville, AL) encoding human chem163S, chem158K, and chem157S lacking the 20-amino acid signal peptide were cloned into pGEX 6P-3 vector (GE Healthcare) and transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The resultant expression plasmids were verified by sequencing.

    Article Title: Studies on the Use of Flagellin as an Immunostimulant and Vaccine Adjuvant in Fish Aquaculture
    Article Snippet: .. Following sequence confirmation, the transformation of BL21 Star (DE3) competent cells (Invitrogen), induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously ( ). .. The monocyte/macrophage-like cell line, RTS-11, from rainbow trout spleen was cultured in Leibovitz (L-15) medium (Invitrogen, United Kingdom) plus 30% fetal calf serum (FCS; Labtech International, United Kingdom) and antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin; Invitrogen, UK) at 20°C, and passaged as described previously ( ).

    Article Title: Rainbow trout CK9, a CCL25-like ancient chemokine that attracts and regulates B cells and macrophages, the main antigen presenting cells in fish
    Article Snippet: The construct (pET-CK9) encodes an identical CK9 mature amino acid sequence with the addition of methionine at the N-terminus and a his-tag (GSGHHHHHHHH) added at the C-terminus for purification. .. Following transformation of the pET-CK9 plasmid into BL21 Star (DE3) competent cells (Invitrogen), the induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ].

    Article Title: Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM+ B Cells in The Absence of Germinal Centers
    Article Snippet: The sequence has been submitted to EMBL/GenBank/DDBJ databases under the accession number FR715329. .. A plasmid was used to transform BL21 Star (DE3) competent cells (Invitrogen) and protein expression was induced by IPTG and purified under denaturing conditions as described previously .

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: .. Expression and Purification of Trout IL-2 Isoforms For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins, and quantification of protein concentration were as described previously ( , , ).

    Article Title: De novo design of a homo-trimeric amantadine-binding protein
    Article Snippet: The resulting expressed protein sequence was as follows: MGSSHHHHHHSSGLVPR/GSHMG//DAQDKLKYLVKQLERALRELKKSLDELERSLEELEKNPSEDALVENNRLNVENNKIIVEVLRIILELAKASAKLA where ‘/' demarks a thrombin cleavage site and ‘//' demarks the beginning of the designed sequence within Rosetta and the numbering of amino acids within this manuscript. .. Constructs were transformed into BL21-Star (DE3) competent cells (Life Technologies).

    Sonication:

    Article Title: Structural Insights into c-Myc-interacting Zinc Finger Protein-1 (Miz-1) Delineate Domains Required for DNA Scanning and Sequence-specific Binding *
    Article Snippet: The construct was transformed in BL21 star (DE3) competent cells (Invitrogen). .. The cell pellet was resuspended in a lysis buffer (700 m m NaCl, 50 m m KH2 PO4 (pH 4.5)), frozen at −80 °C for at least 1 h, thawed in hot water, and then sonicated.

    Affinity Purification:

    Article Title: Arylfluorosulfates Inactivate Intracellular Lipid Binding Protein(s) through Chemoselective SuFEx Reaction with a Binding-site Tyr Residue
    Article Snippet: The pET22b vectors encoding FABP5, FABP4, FABP3 and CRABP2 variants were transformed into BL21 Star (DE3) competent cells (Invitrogen). .. His6 -tagged proteins were affinity purified using Ni-NTA resin (Qiagen) in PBS containing 10 mM imidazole and were eluted with PBS containing 500 mM imidazole.

    Recombinant:

    Article Title: Evolution of Th2 responses: characterization of IL-4/13 in sea bass (Dicentrarchus labrax L.) and studies of expression and biological activity
    Article Snippet: .. Following transformation of the plasmid into BL21 Star (DE3) competent cells (Invitrogen) and the induction of recombinant protein production, a first purification was performed under denaturing conditions. .. Successively we undertook a refolding process, a re-purification under native conditions, a SDS-PAGE analysis of the purified proteins and a quantification of the protein concentration as described in detail previously – .

    Article Title: First in-depth analysis of the novel Th2-type cytokines in salmonid fish reveals distinct patterns of expression and modulation but overlapping bioactivities
    Article Snippet: Expression and purification of trout IL-4/13 isoforms For each protein a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ].

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins, and quantification of protein concentration were as described previously ( , , ).

    Article Title: Studies on the Use of Flagellin as an Immunostimulant and Vaccine Adjuvant in Fish Aquaculture
    Article Snippet: .. Following sequence confirmation, the transformation of BL21 Star (DE3) competent cells (Invitrogen), induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously ( ). .. The monocyte/macrophage-like cell line, RTS-11, from rainbow trout spleen was cultured in Leibovitz (L-15) medium (Invitrogen, United Kingdom) plus 30% fetal calf serum (FCS; Labtech International, United Kingdom) and antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin; Invitrogen, UK) at 20°C, and passaged as described previously ( ).

    Article Title: Rainbow trout CK9, a CCL25-like ancient chemokine that attracts and regulates B cells and macrophages, the main antigen presenting cells in fish
    Article Snippet: .. Following transformation of the pET-CK9 plasmid into BL21 Star (DE3) competent cells (Invitrogen), the induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ]. .. The refolding buffer contained 50 mM Tris-HCl (pH7.5), 10% glycerol, 0.6 M arginine monohydrochloride, 1 M 3-(1-Pyridinio)-1-propanesulfonate (known as NDSB 201 and PPS, Sigma), 0.2% PEG3350 and 5 mM 2-mercaptoethanol.

    Article Title: Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM+ B Cells in The Absence of Germinal Centers
    Article Snippet: Paragraph title: Production of rainbow trout recombinant IL-6 ... A plasmid was used to transform BL21 Star (DE3) competent cells (Invitrogen) and protein expression was induced by IPTG and purified under denaturing conditions as described previously .

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: Expression and Purification of Trout IL-2 Isoforms For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins, and quantification of protein concentration were as described previously ( , , ).

    Molecular Weight:

    Article Title: Evolution of Th2 responses: characterization of IL-4/13 in sea bass (Dicentrarchus labrax L.) and studies of expression and biological activity
    Article Snippet: Thus, the recombinant seabass IL-4/13 isoforms are 133 aa, 131 aa and 140 aa, with a calculated molecular weight of 15.4 kDa, 15.3 kDa and 16.3 kDa, and a theoretical pI of 7.23, 8.58 and 9.14, for IL-4/13-A1, -A2 and -B, respectively. .. Following transformation of the plasmid into BL21 Star (DE3) competent cells (Invitrogen) and the induction of recombinant protein production, a first purification was performed under denaturing conditions.

    Article Title: Rainbow trout CK9, a CCL25-like ancient chemokine that attracts and regulates B cells and macrophages, the main antigen presenting cells in fish
    Article Snippet: Thus, the recombinant trout CK9 is 84 aa, with a calculated molecular weight of 9.61 kDa and a theoretical pI of 9.79. .. Following transformation of the pET-CK9 plasmid into BL21 Star (DE3) competent cells (Invitrogen), the induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ].

    Article Title: Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM+ B Cells in The Absence of Germinal Centers
    Article Snippet: Thus, the rtIL-6 had 211 amino acids, with a predicted molecular weight of 23.8 kDa and pI of 7.27. .. A plasmid was used to transform BL21 Star (DE3) competent cells (Invitrogen) and protein expression was induced by IPTG and purified under denaturing conditions as described previously .

    Mutagenesis:

    Article Title: Studies on the Use of Flagellin as an Immunostimulant and Vaccine Adjuvant in Fish Aquaculture
    Article Snippet: The mutant YRF constructs, YRF-N, YRF-C and YRF-NC, were prepared from pTRI-YRF by PCR using the Q5 high fidelity enzyme (New England Biolabs, United Kingdom) and re-ligation, using primer pairs GCCAGTTCCGCTCATCACCAC/GGAACGGAAGTTACCGTTAACCATC (YRF-N), GCCCATGGTATATCTCCTTTGATTGT/GATAACCGCACGGCAGCCA (YRF-C), and CAAGACTTTAATGCCGTTGAAATCGGT/GTTGAAGCCAAAGGTTTTGACGTATTGA (YRF-NC), respectively. .. Following sequence confirmation, the transformation of BL21 Star (DE3) competent cells (Invitrogen), induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously ( ).

    Size-exclusion Chromatography:

    Article Title: Arylfluorosulfates Inactivate Intracellular Lipid Binding Protein(s) through Chemoselective SuFEx Reaction with a Binding-site Tyr Residue
    Article Snippet: The pET22b vectors encoding FABP5, FABP4, FABP3 and CRABP2 variants were transformed into BL21 Star (DE3) competent cells (Invitrogen). .. Eluted proteins were further purified by size-exclusion chromatography (Superdex 200, GE Healthcare) in PBS.

    Purification:

    Article Title: Evolution of Th2 responses: characterization of IL-4/13 in sea bass (Dicentrarchus labrax L.) and studies of expression and biological activity
    Article Snippet: .. Following transformation of the plasmid into BL21 Star (DE3) competent cells (Invitrogen) and the induction of recombinant protein production, a first purification was performed under denaturing conditions. .. Successively we undertook a refolding process, a re-purification under native conditions, a SDS-PAGE analysis of the purified proteins and a quantification of the protein concentration as described in detail previously – .

    Article Title: Plasticity, dynamics, and inhibition of emerging tetracycline-resistance enzymes
    Article Snippet: Paragraph title: Cloning, expression and purification of tetracycline-inactivating enzymes ... Constructs were transformed into BL21-Star (DE3) competent cells (Life Technologies).

    Article Title: Arylfluorosulfates Inactivate Intracellular Lipid Binding Protein(s) through Chemoselective SuFEx Reaction with a Binding-site Tyr Residue
    Article Snippet: Paragraph title: Protein expression and purification ... The pET22b vectors encoding FABP5, FABP4, FABP3 and CRABP2 variants were transformed into BL21 Star (DE3) competent cells (Invitrogen).

    Article Title: A dual purpose universal influenza vaccine candidate confers protective immunity against anthrax
    Article Snippet: Paragraph title: Vaccine candidate design, production and purification ... All expression plasmids were transformed in BL21 Star™ (DE3) competent cells (Life Technologies).

    Article Title: First in-depth analysis of the novel Th2-type cytokines in salmonid fish reveals distinct patterns of expression and modulation but overlapping bioactivities
    Article Snippet: .. Expression and purification of trout IL-4/13 isoforms For each protein a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ].

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: Paragraph title: Expression and Purification of Trout IL-2 Isoforms ... For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen).

    Article Title: Studies on the Use of Flagellin as an Immunostimulant and Vaccine Adjuvant in Fish Aquaculture
    Article Snippet: .. Following sequence confirmation, the transformation of BL21 Star (DE3) competent cells (Invitrogen), induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously ( ). .. The monocyte/macrophage-like cell line, RTS-11, from rainbow trout spleen was cultured in Leibovitz (L-15) medium (Invitrogen, United Kingdom) plus 30% fetal calf serum (FCS; Labtech International, United Kingdom) and antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin; Invitrogen, UK) at 20°C, and passaged as described previously ( ).

    Article Title: Rainbow trout CK9, a CCL25-like ancient chemokine that attracts and regulates B cells and macrophages, the main antigen presenting cells in fish
    Article Snippet: .. Following transformation of the pET-CK9 plasmid into BL21 Star (DE3) competent cells (Invitrogen), the induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ]. .. The refolding buffer contained 50 mM Tris-HCl (pH7.5), 10% glycerol, 0.6 M arginine monohydrochloride, 1 M 3-(1-Pyridinio)-1-propanesulfonate (known as NDSB 201 and PPS, Sigma), 0.2% PEG3350 and 5 mM 2-mercaptoethanol.

    Article Title: Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM+ B Cells in The Absence of Germinal Centers
    Article Snippet: .. A plasmid was used to transform BL21 Star (DE3) competent cells (Invitrogen) and protein expression was induced by IPTG and purified under denaturing conditions as described previously . .. The purified, denatured rtIL-6 was refolded in refolding buffer containing 50 mM Tris–HCl, pH8.0, 0.5 M arginine, 0.5% Triton-100 and 5 mM β-mercaptoethanol at 4 °C for 2 days.

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: .. Expression and Purification of Trout IL-2 Isoforms For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins, and quantification of protein concentration were as described previously ( , , ).

    Article Title: De novo design of a homo-trimeric amantadine-binding protein
    Article Snippet: Paragraph title: Cloning, protein expression and purification ... Constructs were transformed into BL21-Star (DE3) competent cells (Life Technologies).

    Polymerase Chain Reaction:

    Article Title: Structural Insights into c-Myc-interacting Zinc Finger Protein-1 (Miz-1) Delineate Domains Required for DNA Scanning and Sequence-specific Binding *
    Article Snippet: The PCR product was inserted into pDrive (Qiagen), digested by NdeI and BamHI, and inserted in pET-3a (Novagen) by the same restriction sites. .. The construct was transformed in BL21 star (DE3) competent cells (Invitrogen).

    Article Title: Studies on the Use of Flagellin as an Immunostimulant and Vaccine Adjuvant in Fish Aquaculture
    Article Snippet: The mutant YRF constructs, YRF-N, YRF-C and YRF-NC, were prepared from pTRI-YRF by PCR using the Q5 high fidelity enzyme (New England Biolabs, United Kingdom) and re-ligation, using primer pairs GCCAGTTCCGCTCATCACCAC/GGAACGGAAGTTACCGTTAACCATC (YRF-N), GCCCATGGTATATCTCCTTTGATTGT/GATAACCGCACGGCAGCCA (YRF-C), and CAAGACTTTAATGCCGTTGAAATCGGT/GTTGAAGCCAAAGGTTTTGACGTATTGA (YRF-NC), respectively. .. Following sequence confirmation, the transformation of BL21 Star (DE3) competent cells (Invitrogen), induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously ( ).

    Affinity Chromatography:

    Article Title: A dual purpose universal influenza vaccine candidate confers protective immunity against anthrax
    Article Snippet: All expression plasmids were transformed in BL21 Star™ (DE3) competent cells (Life Technologies). .. The expressed proteins contained 6×His tags and were purified by affinity chromatography using Ni‐NTA agarose beads.

    SDS Page:

    Article Title: Evolution of Th2 responses: characterization of IL-4/13 in sea bass (Dicentrarchus labrax L.) and studies of expression and biological activity
    Article Snippet: Following transformation of the plasmid into BL21 Star (DE3) competent cells (Invitrogen) and the induction of recombinant protein production, a first purification was performed under denaturing conditions. .. Successively we undertook a refolding process, a re-purification under native conditions, a SDS-PAGE analysis of the purified proteins and a quantification of the protein concentration as described in detail previously – .

    Article Title: First in-depth analysis of the novel Th2-type cytokines in salmonid fish reveals distinct patterns of expression and modulation but overlapping bioactivities
    Article Snippet: Expression and purification of trout IL-4/13 isoforms For each protein a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ].

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins, and quantification of protein concentration were as described previously ( , , ).

    Article Title: Studies on the Use of Flagellin as an Immunostimulant and Vaccine Adjuvant in Fish Aquaculture
    Article Snippet: .. Following sequence confirmation, the transformation of BL21 Star (DE3) competent cells (Invitrogen), induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously ( ). .. The monocyte/macrophage-like cell line, RTS-11, from rainbow trout spleen was cultured in Leibovitz (L-15) medium (Invitrogen, United Kingdom) plus 30% fetal calf serum (FCS; Labtech International, United Kingdom) and antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin; Invitrogen, UK) at 20°C, and passaged as described previously ( ).

    Article Title: Rainbow trout CK9, a CCL25-like ancient chemokine that attracts and regulates B cells and macrophages, the main antigen presenting cells in fish
    Article Snippet: .. Following transformation of the pET-CK9 plasmid into BL21 Star (DE3) competent cells (Invitrogen), the induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ]. .. The refolding buffer contained 50 mM Tris-HCl (pH7.5), 10% glycerol, 0.6 M arginine monohydrochloride, 1 M 3-(1-Pyridinio)-1-propanesulfonate (known as NDSB 201 and PPS, Sigma), 0.2% PEG3350 and 5 mM 2-mercaptoethanol.

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: Expression and Purification of Trout IL-2 Isoforms For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins, and quantification of protein concentration were as described previously ( , , ).

    Plasmid Preparation:

    Article Title: Evolution of Th2 responses: characterization of IL-4/13 in sea bass (Dicentrarchus labrax L.) and studies of expression and biological activity
    Article Snippet: .. Following transformation of the plasmid into BL21 Star (DE3) competent cells (Invitrogen) and the induction of recombinant protein production, a first purification was performed under denaturing conditions. .. Successively we undertook a refolding process, a re-purification under native conditions, a SDS-PAGE analysis of the purified proteins and a quantification of the protein concentration as described in detail previously – .

    Article Title: Plasticity, dynamics, and inhibition of emerging tetracycline-resistance enzymes
    Article Snippet: Cloning, expression and purification of tetracycline-inactivating enzymes All genes encoding tetracycline-inactivating enzymes were cloned into the pET28b(+) vector (Novagen) at BamHI and NdeI restriction sites. .. Constructs were transformed into BL21-Star (DE3) competent cells (Life Technologies).

    Article Title: A dual purpose universal influenza vaccine candidate confers protective immunity against anthrax
    Article Snippet: For EFn‐M2, the EFn sequence was cloned upstream of the full‐length M2 sequence in the pET200/D‐TOPO plasmid. .. All expression plasmids were transformed in BL21 Star™ (DE3) competent cells (Life Technologies).

    Article Title: First in-depth analysis of the novel Th2-type cytokines in salmonid fish reveals distinct patterns of expression and modulation but overlapping bioactivities
    Article Snippet: .. Expression and purification of trout IL-4/13 isoforms For each protein a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ].

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: .. For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins, and quantification of protein concentration were as described previously ( , , ).

    Article Title: Chemerin158K Protein Is the Dominant Chemerin Isoform in Synovial and Cerebrospinal Fluids but Not in Plasma *
    Article Snippet: .. cDNA (Open Biosystems, Huntsville, AL) encoding human chem163S, chem158K, and chem157S lacking the 20-amino acid signal peptide were cloned into pGEX 6P-3 vector (GE Healthcare) and transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The resultant expression plasmids were verified by sequencing.

    Article Title: Rainbow trout CK9, a CCL25-like ancient chemokine that attracts and regulates B cells and macrophages, the main antigen presenting cells in fish
    Article Snippet: .. Following transformation of the pET-CK9 plasmid into BL21 Star (DE3) competent cells (Invitrogen), the induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ]. .. The refolding buffer contained 50 mM Tris-HCl (pH7.5), 10% glycerol, 0.6 M arginine monohydrochloride, 1 M 3-(1-Pyridinio)-1-propanesulfonate (known as NDSB 201 and PPS, Sigma), 0.2% PEG3350 and 5 mM 2-mercaptoethanol.

    Article Title: Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM+ B Cells in The Absence of Germinal Centers
    Article Snippet: .. A plasmid was used to transform BL21 Star (DE3) competent cells (Invitrogen) and protein expression was induced by IPTG and purified under denaturing conditions as described previously . .. The purified, denatured rtIL-6 was refolded in refolding buffer containing 50 mM Tris–HCl, pH8.0, 0.5 M arginine, 0.5% Triton-100 and 5 mM β-mercaptoethanol at 4 °C for 2 days.

    Article Title: Interleukin (IL)-2 Is a Key Regulator of T Helper 1 and T Helper 2 Cytokine Expression in Fish: Functional Characterization of Two Divergent IL2 Paralogs in Salmonids
    Article Snippet: .. Expression and Purification of Trout IL-2 Isoforms For each protein, a sequence confirmed plasmid was transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins, and quantification of protein concentration were as described previously ( , , ).

    Article Title: De novo design of a homo-trimeric amantadine-binding protein
    Article Snippet: Cloning, protein expression and purification ABP was cloned into the pET28b(+) vector at NdeI and XhoI restriction sites. .. Constructs were transformed into BL21-Star (DE3) competent cells (Life Technologies).

    Positron Emission Tomography:

    Article Title: Structural Insights into c-Myc-interacting Zinc Finger Protein-1 (Miz-1) Delineate Domains Required for DNA Scanning and Sequence-specific Binding *
    Article Snippet: The PCR product was inserted into pDrive (Qiagen), digested by NdeI and BamHI, and inserted in pET-3a (Novagen) by the same restriction sites. .. The construct was transformed in BL21 star (DE3) competent cells (Invitrogen).

    Article Title: Rainbow trout CK9, a CCL25-like ancient chemokine that attracts and regulates B cells and macrophages, the main antigen presenting cells in fish
    Article Snippet: .. Following transformation of the pET-CK9 plasmid into BL21 Star (DE3) competent cells (Invitrogen), the induction of recombinant protein production, purification under denaturing conditions, refolding, re-purification under native conditions, SDS-PAGE analysis of proteins and quantification of protein concentration were as described previously [ , , ]. .. The refolding buffer contained 50 mM Tris-HCl (pH7.5), 10% glycerol, 0.6 M arginine monohydrochloride, 1 M 3-(1-Pyridinio)-1-propanesulfonate (known as NDSB 201 and PPS, Sigma), 0.2% PEG3350 and 5 mM 2-mercaptoethanol.

    Produced:

    Article Title: Chemerin158K Protein Is the Dominant Chemerin Isoform in Synovial and Cerebrospinal Fluids but Not in Plasma *
    Article Snippet: cDNA (Open Biosystems, Huntsville, AL) encoding human chem163S, chem158K, and chem157S lacking the 20-amino acid signal peptide were cloned into pGEX 6P-3 vector (GE Healthcare) and transformed into BL21 Star (DE3) competent cells (Invitrogen). .. The proteins were produced in 2× YT medium at 37 °C for 4 h. Cells were pelleted before suspension in STE buffer (50 m m Tris-HCl, 150 m m NaCl, 1 m m EDTA, pH 7.5) containing 1 m m DTT, 5 units/ml rLysozyme (Novagen, Madison, WI), benzonase nuclease, and Complete protease inhibitor (Roche Applied Science).

    Concentration Assay:

    Article Title: De novo design of a homo-trimeric amantadine-binding protein
    Article Snippet: Constructs were transformed into BL21-Star (DE3) competent cells (Life Technologies). .. Cells harboring the plasmid were grown at 37°C in Terrific Broth II medium containing a final concentration of 0.05 mg/ml kanamycin.

    CTG Assay:

    Article Title: Structural Insights into c-Myc-interacting Zinc Finger Protein-1 (Miz-1) Delineate Domains Required for DNA Scanning and Sequence-specific Binding *
    Article Snippet: The cDNA of ZFs 1–4 of Miz-1 (Miz1–4, residues 304–414) was generated by PCR from the complete cDNA of Miz-1 using primers F (5′-CAT ATG GTC ATC CAC AAG TGC GAG GAC TGT GG-3′) and R (5′-GGA TCC CTA GCC GCT GTG CAC CAG CTG GTG-3′). .. The construct was transformed in BL21 star (DE3) competent cells (Invitrogen).

    Lysis:

    Article Title: Structural Insights into c-Myc-interacting Zinc Finger Protein-1 (Miz-1) Delineate Domains Required for DNA Scanning and Sequence-specific Binding *
    Article Snippet: The construct was transformed in BL21 star (DE3) competent cells (Invitrogen). .. The cell pellet was resuspended in a lysis buffer (700 m m NaCl, 50 m m KH2 PO4 (pH 4.5)), frozen at −80 °C for at least 1 h, thawed in hot water, and then sonicated.

    Fluorescence In Situ Hybridization:

    Article Title: Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM+ B Cells in The Absence of Germinal Centers
    Article Snippet: Production of rainbow trout recombinant IL-6 The sequence encoding the mature peptide of trout IL-6 was amplified from spleen cDNA prepared from Aeromonas salmonicida infected fish and cloned to the pET/Duet-1 vector (Novagen, UK). .. A plasmid was used to transform BL21 Star (DE3) competent cells (Invitrogen) and protein expression was induced by IPTG and purified under denaturing conditions as described previously .

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