shuffle t7 (New England Biolabs)

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SHuffle T7 Competent E coli

Description:

SHuffle T7 Competent E coli 12x0 05 ml

Catalog Number:

c3026j

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228

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12x0 05 ml

Category:

Competent Bacteria

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New England Biolabs shuffle t7

SHuffle T7 Competent E coli 12x0 05 ml
https://www.bioz.com/result/shuffle t7/product/New England Biolabs
Average 95 stars, based on 43 article reviews
Price from $9.99 to $1999.99

shuffle t7 - by Bioz Stars, 2020-02

95/100 stars

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Clone Assay:

Article Title: Bifunctional Chloroplastic DJ-1B from Arabidopsis thaliana is an Oxidation-Robust Holdase and a Glyoxalase Sensitive to H2O2
Article Snippet: Paragraph title: 2.1. Cloning and Purification of Recombinant AtDJ-1B ... This expression vector was transformed into SHuffle® T7 competent E. coli (New England Biolabs, Ipswitch, MA, USA) and plated on agar to obtain a single colony, which was grown overnight at 30 °C in Luria-Bertani Broth (LB) supplemented with 100 μg/mL ampicillin.

Article Title: Bacterial Expression of Human Butyrylcholinesterase as a Tool for Nerve Agent Bioscavengers Development
Article Snippet: Expression and Purification For the initial expression screening, the different expression vectors (pThioHis-hBChE-1 to pThioHis-hBChE-7) were independently transformed into SHuffle® T7 Competent E. coli (New England Biolabs, Evry, France). .. Clones of each construct were cultured overnight in 5 mL of LB (Sigma Aldrich, Saint-Quentin-Fallavier, France), supplemented by 100 µg mL−1 ampicillin (Sigma Aldrich, Saint-Quentin-Fallavier, France) at 37 °C, and protein synthesis was induced by addition of 1 mM IPTG (Acros Organics) for 4 h at 37 °C.

Article Title: Efficient biosynthesis of heterodimeric C3-aryl pyrroloindoline alkaloids
Article Snippet: .. The B type strains are often used for protein expression, while K type strains are mostly used for DNA cloning but also for protein expression, such as Shuffle T7 competent E. coli from New England Biolabs. ..

Article Title: The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall
Article Snippet: .. Cloning was done using E. coli DH5α and selection with 30 µg/ml kanamycin; the final expression vector was transformed into SHuffle T7 competent E. coli cells (New England Biolabs). .. LB medium containing selection antibiotic was inoculated with the expression strain and shaken at 200 rpm and 30°C.

Article Title: A genetically encoded fluorescent temperature sensor derived from the photoactive Orange Carotenoid Protein
Article Snippet: Paragraph title: Cloning, protein expression, and purification ... To produce chimeric proteins, SHuffle® T7 Competent E. coli (NEB, USA) cells were transformed by the resultant pQE81L-TagRFP-OCP and pQE81L-OCP-TagGFP plasmid constructs and incubated in LB medium overnight at 37 °C.

Centrifugation:

Article Title: Escherichia coli-based production of recombinant ovine angiotensinogen and its characterization as a renin substrate
Article Snippet: Expression of recombinant oANG in E. coli The pET-11a-oANG plasmid was used to transform six different E. coli host strains: BL21(DE3), Rosetta 2(DE3), Tuner(DE3), HMS174(DE3) (Novagen), SHuffle T7 Express Competent E. coli (SHB), and SHuffle T7 Competent E. coli (SHK) (New England Biolabs, Herts, UK). .. Cells were collected by centrifugation (2,200 × g , 10 min, 4 °C), resuspended in phosphate-buffered saline (10 mM Na2 HPO4 , 1.8 mM KH2 PO4 , 140 mM NaCl, and 2.7 mM KCl, pH 7.3), and disrupted by sonication.

Article Title: Bacterial Expression of Human Butyrylcholinesterase as a Tool for Nerve Agent Bioscavengers Development
Article Snippet: Expression and Purification For the initial expression screening, the different expression vectors (pThioHis-hBChE-1 to pThioHis-hBChE-7) were independently transformed into SHuffle® T7 Competent E. coli (New England Biolabs, Evry, France). .. Total soluble extracts were prepared by pulsed sonication on ice for 3 times 5 min using a S2 micro-probe (2 mm) equipped UP200S sonicator (Hielscher, Teltow, Germany) and centrifugation at 20,000× g for 20 min at 4 °C.

Article Title: The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall
Article Snippet: Cloning was done using E. coli DH5α and selection with 30 µg/ml kanamycin; the final expression vector was transformed into SHuffle T7 competent E. coli cells (New England Biolabs). .. Cells were collected after 4 h (centrifugation, 30 min at 4,000 × g ) and suspended in 50 mM Tris-Cl (pH 8.0) containing 200 µg/ml lysozyme (Sigma-Aldrich).

Article Title: Plasmodium falciparum MSP3 Exists in a Complex on the Merozoite Surface and Generates Antibody Response during Natural Infection
Article Snippet: The large N-terminal fragment of MSP3N in a pET-28a expression construct was transformed into SHuffle T7 competent E. coli cells (NEB) for expression; the cells were induced with 0.5 mM isopropyl- d -thiogalactopyranoside (IPTG) (Sigma Chemical Co.) for 8 h at 30°C. .. The soluble and insoluble fractions were separated by centrifugation at 12,000 × g for 30 min and analyzed by SDS-PAGE.

Amplification:

Article Title: A genetically encoded fluorescent temperature sensor derived from the photoactive Orange Carotenoid Protein
Article Snippet: The PCR product of TagGFP was utilized for overlapping PCR with the amplified sequence of Synechocystis OCP; the resulting OCP-TagGFP PCR product was cloned into the pQE81L expression vector utilizing BamHI and NotI restriction sites (New England Biolabs, USA). .. To produce chimeric proteins, SHuffle® T7 Competent E. coli (NEB, USA) cells were transformed by the resultant pQE81L-TagRFP-OCP and pQE81L-OCP-TagGFP plasmid constructs and incubated in LB medium overnight at 37 °C.

Synthesized:

Article Title: The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall
Article Snippet: The gene was synthesized with flanking NdeI and XhoI restriction sites (GeneArt; Life Technologies, Inc.). .. Cloning was done using E. coli DH5α and selection with 30 µg/ml kanamycin; the final expression vector was transformed into SHuffle T7 competent E. coli cells (New England Biolabs).

Construct:

Article Title: Overlapping Specificity of Duplicated Human Pancreatic Elastase 3 Isoforms and Archetypal Porcine Elastase 1 Provides Clues to Evolution of Digestive Enzymes *
Article Snippet: The recombinant SGPI-2 variants were expressed into the cytoplasm of E. coli SHuffle T7 cells (C3026, New England Biolabs) fused to the C terminus of the E. coli protein-disulfide isomerase DsbC following the strategy of Nozach et al. ( ). .. The construct also incorporated an N-terminal His tag and a TEV proteinase cleavage site between the inhibitor and DsbC.

Article Title: A genetically encoded fluorescent temperature sensor derived from the photoactive Orange Carotenoid Protein
Article Snippet: .. To produce chimeric proteins, SHuffle® T7 Competent E. coli (NEB, USA) cells were transformed by the resultant pQE81L-TagRFP-OCP and pQE81L-OCP-TagGFP plasmid constructs and incubated in LB medium overnight at 37 °C. .. At an OD550 of 0.6, protein expression was induced with 1 mM IPTG and continued for 6 h at 30 °C.

Article Title: Nuclear Import of Arabidopsis Poly(ADP-Ribose) Polymerase 2 Is Mediated by Importin-α and a Nuclear Localization Sequence Located Between the Predicted SAP Domains
Article Snippet: .. Protein Expression and Purification For protein expression in E. coli , pOPINS3C carrying His6:SUMO:SAP and pOPINE carrying the α-GFP-nanobody:Halo:His6 construct (Addgene plasmid #111090) were transformed into SHuffle® T7 Competent E. coli cells (New England Biolabs). .. The His6:ΔIBB-IMPORTIN-α2 protein was expressed from pOPINF in SoluBL21 cells (Genlantis).

Article Title: Plasmodium falciparum MSP3 Exists in a Complex on the Merozoite Surface and Generates Antibody Response during Natural Infection
Article Snippet: .. The large N-terminal fragment of MSP3N in a pET-28a expression construct was transformed into SHuffle T7 competent E. coli cells (NEB) for expression; the cells were induced with 0.5 mM isopropyl- d -thiogalactopyranoside (IPTG) (Sigma Chemical Co.) for 8 h at 30°C. .. The induced cell pellet was disrupted by sonication in Tris-NaCl lysis buffer (0.05 M Tris, pH 8.0, and 0.15 M NaCl, 0.01 M dithiothreitol [DTT], 1 mM phenylmethylsulfonyl fluoride [PMSF], 1% Triton X-100) with 9-s pulses at 9-s intervals 10 times using a miniprobe.

Article Title: Nuclear Import of Arabidopsis Poly(ADP-Ribose) Polymerase 2 Is Mediated by Importin-α and a Nuclear Localization Sequence Located Between the Predicted SAP Domains
Article Snippet: .. For protein expression in E. coli , pOPINS3C carrying His6:SUMO:SAP and pOPINE carrying the α-GFP-nanobody:Halo:His6 construct (Addgene plasmid #111090) were transformed into SHuffle® T7 Competent E. coli cells (New England Biolabs). .. The His6:ΔIBB-IMPORTIN-α2 protein was expressed from pOPINF in SoluBL21 cells (Genlantis).

Incubation:

Article Title: Biochemical characterization of Plasmodium falciparum parasite specific helicase 1 (PfPSH1)
Article Snippet: Expression and purification of recombinant proteins To obtain recombinant PfPSH1C, PfPSH1C‐pET28a clone was transformed in SHuffle T7 competent E. coli cells from New England Biolabs (Ipswich, MA, USA). .. For overexpression, PfPSH1C primary culture was incubated overnight and secondary culture was inoculated with 4% of overnight grown primary inoculum.

Solubility:

Article Title: Escherichia coli-based production of recombinant ovine angiotensinogen and its characterization as a renin substrate
Article Snippet: Expression of recombinant oANG in E. coli The pET-11a-oANG plasmid was used to transform six different E. coli host strains: BL21(DE3), Rosetta 2(DE3), Tuner(DE3), HMS174(DE3) (Novagen), SHuffle T7 Express Competent E. coli (SHB), and SHuffle T7 Competent E. coli (SHK) (New England Biolabs, Herts, UK). .. After the transformed cells were grown at 37 °C to mid-log phase (OD600 approximately 0.6), IPTG was added to the culture medium and protein expression was induced for 3 h. IPTG concentrations and expression temperatures were varied in an attempt to improve the solubility of oANG.

Expressing:

Article Title: Bifunctional Chloroplastic DJ-1B from Arabidopsis thaliana is an Oxidation-Robust Holdase and a Glyoxalase Sensitive to H2O2
Article Snippet: .. This expression vector was transformed into SHuffle® T7 competent E. coli (New England Biolabs, Ipswitch, MA, USA) and plated on agar to obtain a single colony, which was grown overnight at 30 °C in Luria-Bertani Broth (LB) supplemented with 100 μg/mL ampicillin. ..

Article Title: Escherichia coli-based production of recombinant ovine angiotensinogen and its characterization as a renin substrate
Article Snippet: .. Expression of recombinant oANG in E. coli The pET-11a-oANG plasmid was used to transform six different E. coli host strains: BL21(DE3), Rosetta 2(DE3), Tuner(DE3), HMS174(DE3) (Novagen), SHuffle T7 Express Competent E. coli (SHB), and SHuffle T7 Competent E. coli (SHK) (New England Biolabs, Herts, UK). .. Ten randomly selected colonies were suspended in 0.05 mL of LB medium (Nacalai, Kyoto, Japan), and 0.02 mL of the suspensions were added to 2 mL of LB medium supplemented with either 50 μg/mL ampicillin [all host strains except for Rosetta 2(DE3)] or 50 μg/mL ampicillin and 25 μg/mL chloramphenicol [Rosetta 2(DE3)].

Article Title: Bacterial Expression of Human Butyrylcholinesterase as a Tool for Nerve Agent Bioscavengers Development
Article Snippet: .. Expression and Purification For the initial expression screening, the different expression vectors (pThioHis-hBChE-1 to pThioHis-hBChE-7) were independently transformed into SHuffle® T7 Competent E. coli (New England Biolabs, Evry, France). .. Clones of each construct were cultured overnight in 5 mL of LB (Sigma Aldrich, Saint-Quentin-Fallavier, France), supplemented by 100 µg mL−1 ampicillin (Sigma Aldrich, Saint-Quentin-Fallavier, France) at 37 °C, and protein synthesis was induced by addition of 1 mM IPTG (Acros Organics) for 4 h at 37 °C.

Article Title: Overlapping Specificity of Duplicated Human Pancreatic Elastase 3 Isoforms and Archetypal Porcine Elastase 1 Provides Clues to Evolution of Digestive Enzymes *
Article Snippet: Paragraph title: Expression and Purification of SGPI-2 Variants ... The recombinant SGPI-2 variants were expressed into the cytoplasm of E. coli SHuffle T7 cells (C3026, New England Biolabs) fused to the C terminus of the E. coli protein-disulfide isomerase DsbC following the strategy of Nozach et al. ( ).

Article Title: Biochemical characterization of Plasmodium falciparum parasite specific helicase 1 (PfPSH1)
Article Snippet: .. Expression and purification of recombinant proteins To obtain recombinant PfPSH1C, PfPSH1C‐pET28a clone was transformed in SHuffle T7 competent E. coli cells from New England Biolabs (Ipswich, MA, USA). .. For overexpression, PfPSH1C primary culture was incubated overnight and secondary culture was inoculated with 4% of overnight grown primary inoculum.

Article Title: The mechanism of GM-CSF inhibition by human GM-CSF auto-antibodies suggests novel therapeutic opportunities
Article Snippet: .. Human GM-CSF (residues A1-E127 of the mature peptide) was expressed in E. coli and purified by anion exchange chromatography and reversed phase HPLC., Recombinant human Fab F1 was expressed in E. coli SHuffle T7 cells (New England Biolab, Cat. C3026J) using the pET-Duet (Millipore-Novagen, Cat. 71146–3) expression plasmid containing the genes for the light and heavy chains of F1 Fab. ..

Article Title: Nuclear Import of Arabidopsis Poly(ADP-Ribose) Polymerase 2 Is Mediated by Importin-α and a Nuclear Localization Sequence Located Between the Predicted SAP Domains
Article Snippet: .. For protein expression in E. coli , pOPINS3C carrying His6:SUMO:SAP and pOPINE carrying the α-GFP-nanobody:Halo:His6 construct (Addgene plasmid #111090) were transformed into SHuffle® T7 Competent E. coli cells (New England Biolabs). .. The His6:ΔIBB-IMPORTIN-α2 protein was expressed from pOPINF in SoluBL21 cells (Genlantis).

Transformation Assay:

Article Title: Bacterial Expression of Human Butyrylcholinesterase as a Tool for Nerve Agent Bioscavengers Development
Article Snippet: .. For large scale production and purification, p8HisThioTEV-hBChE-7 vector was transformed into SHuffle® T7 Competent E. coli and 3 L of LB were prepared after IPTG induction at OD600nm = 0.8 and overnight culture at 37 °C. .. Bacterial pellet was suspended in 150 mL of 20 mM HEPES pH 7.5, 0.3 M NaCl, 15 mM Imidazole and sonicated on ice for 5 times 3 min with 0.5 s pulses using a 12.5 mm probe equipped Sonic Ruptor 400 (Omni, Kennesaw, GA, USA).

Article Title: Escherichia coli-based production of recombinant ovine angiotensinogen and its characterization as a renin substrate
Article Snippet: Expression of recombinant oANG in E. coli The pET-11a-oANG plasmid was used to transform six different E. coli host strains: BL21(DE3), Rosetta 2(DE3), Tuner(DE3), HMS174(DE3) (Novagen), SHuffle T7 Express Competent E. coli (SHB), and SHuffle T7 Competent E. coli (SHK) (New England Biolabs, Herts, UK). .. After the transformed cells were grown at 37 °C to mid-log phase (OD600 approximately 0.6), IPTG was added to the culture medium and protein expression was induced for 3 h. IPTG concentrations and expression temperatures were varied in an attempt to improve the solubility of oANG.

Article Title: Nuclear Import of Arabidopsis Poly(ADP-Ribose) Polymerase 2 Is Mediated by Importin-α and a Nuclear Localization Sequence Located Between the Predicted SAP Domains
Article Snippet: .. For protein expression in E. coli , pOPINS3C carrying His6:SUMO:SAP and pOPINE carrying the α-GFP-nanobody:Halo:His6 construct (Addgene plasmid #111090) were transformed into SHuffle® T7 Competent E. coli cells (New England Biolabs). .. The His6:ΔIBB-IMPORTIN-α2 protein was expressed from pOPINF in SoluBL21 cells (Genlantis).

Over Expression:

Derivative Assay:

Article Title: Efficient biosynthesis of heterodimeric C3-aryl pyrroloindoline alkaloids
Article Snippet: As cells are safe upon the individual expression of Fd, FdR, and NascB, the toxicity effect must be derived from the activated NascB in the presence of both Fd and FdR. .. The B type strains are often used for protein expression, while K type strains are mostly used for DNA cloning but also for protein expression, such as Shuffle T7 competent E. coli from New England Biolabs.

Article Title: A genetically encoded fluorescent temperature sensor derived from the photoactive Orange Carotenoid Protein
Article Snippet: The 6xHis-tag derived from the pQE81L vector construct was identical in both types of chimeras. .. To produce chimeric proteins, SHuffle® T7 Competent E. coli (NEB, USA) cells were transformed by the resultant pQE81L-TagRFP-OCP and pQE81L-OCP-TagGFP plasmid constructs and incubated in LB medium overnight at 37 °C.

High Performance Liquid Chromatography:

Chromatography:

Article Title: Overlapping Specificity of Duplicated Human Pancreatic Elastase 3 Isoforms and Archetypal Porcine Elastase 1 Provides Clues to Evolution of Digestive Enzymes *
Article Snippet: The recombinant SGPI-2 variants were expressed into the cytoplasm of E. coli SHuffle T7 cells (C3026, New England Biolabs) fused to the C terminus of the E. coli protein-disulfide isomerase DsbC following the strategy of Nozach et al. ( ). .. The fusion protein was purified by nickel-affinity chromatography and processed by His-tagged TEV proteinase.

Article Title: A genetically encoded fluorescent temperature sensor derived from the photoactive Orange Carotenoid Protein
Article Snippet: To produce chimeric proteins, SHuffle® T7 Competent E. coli (NEB, USA) cells were transformed by the resultant pQE81L-TagRFP-OCP and pQE81L-OCP-TagGFP plasmid constructs and incubated in LB medium overnight at 37 °C. .. All proteins were purified by immobilized metal-affinity and anion exchange chromatography to electrophoretic homogeneity and stored at 4 °C in the presence of 2 mM sodium azide.

Protease Inhibitor:

Article Title: Biochemical characterization of Plasmodium falciparum parasite specific helicase 1 (PfPSH1)
Article Snippet: Expression and purification of recombinant proteins To obtain recombinant PfPSH1C, PfPSH1C‐pET28a clone was transformed in SHuffle T7 competent E. coli cells from New England Biolabs (Ipswich, MA, USA). .. The buffer (50 mm Tris/HCl, 500 mm NaCl, 0.05% Tween‐20, 0.1% Triton X‐100, and 0.5% CHAPS) and the protease inhibitor cocktail from Roche (Sigma, St. Louis, MO, USA) were used for sonicating the lysate.

Cell Culture:

Polymerase Chain Reaction:

Article Title: Bifunctional Chloroplastic DJ-1B from Arabidopsis thaliana is an Oxidation-Robust Holdase and a Glyoxalase Sensitive to H2O2
Article Snippet: An N-terminal Tobacco Etch Virus (TEV)-cleavage site was introduced into a codon-optimized Arabidopsis AtDJ-1B open reading frame by polymerase chain reaction (PCR), before subcloning into a pDEST15 expression vector with an N-terminal glutathione-S-transferase (GST)-tag (Gateway technology, Thermo Fisher Scientific, Waltham, MA, USA)). .. This expression vector was transformed into SHuffle® T7 competent E. coli (New England Biolabs, Ipswitch, MA, USA) and plated on agar to obtain a single colony, which was grown overnight at 30 °C in Luria-Bertani Broth (LB) supplemented with 100 μg/mL ampicillin.

Article Title: Overlapping Specificity of Duplicated Human Pancreatic Elastase 3 Isoforms and Archetypal Porcine Elastase 1 Provides Clues to Evolution of Digestive Enzymes *
Article Snippet: Based on the sequence pattern of the inhibitor binding loops selected against the human elastases, 12 different inhibitor variants were designed and created using PCR-based mutagenesis ( ). .. The recombinant SGPI-2 variants were expressed into the cytoplasm of E. coli SHuffle T7 cells (C3026, New England Biolabs) fused to the C terminus of the E. coli protein-disulfide isomerase DsbC following the strategy of Nozach et al. ( ).

Article Title: Bifunctional Chloroplastic DJ-1B from Arabidopsis thaliana is an Oxidation-Robust Holdase and a Glyoxalase Sensitive to H2O2
Article Snippet: Cloning and Purification of Recombinant AtDJ-1B An N-terminal Tobacco Etch Virus (TEV)-cleavage site was introduced into a codon-optimized Arabidopsis AtDJ-1B open reading frame by polymerase chain reaction (PCR), before subcloning into a pDEST15 expression vector with an N-terminal glutathione-S-transferase (GST)-tag (Gateway technology, Thermo Fisher Scientific, Waltham, MA, USA)). .. This expression vector was transformed into SHuffle® T7 competent E. coli (New England Biolabs, Ipswitch, MA, USA) and plated on agar to obtain a single colony, which was grown overnight at 30 °C in Luria-Bertani Broth (LB) supplemented with 100 μg/mL ampicillin.

Sonication:

Article Title: Bacterial Expression of Human Butyrylcholinesterase as a Tool for Nerve Agent Bioscavengers Development
Article Snippet: Expression and Purification For the initial expression screening, the different expression vectors (pThioHis-hBChE-1 to pThioHis-hBChE-7) were independently transformed into SHuffle® T7 Competent E. coli (New England Biolabs, Evry, France). .. Total soluble extracts were prepared by pulsed sonication on ice for 3 times 5 min using a S2 micro-probe (2 mm) equipped UP200S sonicator (Hielscher, Teltow, Germany) and centrifugation at 20,000× g for 20 min at 4 °C.

Article Title: Plasmodium falciparum MSP3 Exists in a Complex on the Merozoite Surface and Generates Antibody Response during Natural Infection
Article Snippet: The large N-terminal fragment of MSP3N in a pET-28a expression construct was transformed into SHuffle T7 competent E. coli cells (NEB) for expression; the cells were induced with 0.5 mM isopropyl- d -thiogalactopyranoside (IPTG) (Sigma Chemical Co.) for 8 h at 30°C. .. The induced cell pellet was disrupted by sonication in Tris-NaCl lysis buffer (0.05 M Tris, pH 8.0, and 0.15 M NaCl, 0.01 M dithiothreitol [DTT], 1 mM phenylmethylsulfonyl fluoride [PMSF], 1% Triton X-100) with 9-s pulses at 9-s intervals 10 times using a miniprobe.

Binding Assay:

Article Title: Bifunctional Chloroplastic DJ-1B from Arabidopsis thaliana is an Oxidation-Robust Holdase and a Glyoxalase Sensitive to H2O2
Article Snippet: This expression vector was transformed into SHuffle® T7 competent E. coli (New England Biolabs, Ipswitch, MA, USA) and plated on agar to obtain a single colony, which was grown overnight at 30 °C in Luria-Bertani Broth (LB) supplemented with 100 μg/mL ampicillin. .. The supernatant was passed through a 0.45-μm filter and loaded onto Glutathione (GSH) Sepharose High Performance resin (GE Healthcare Europe, Diegem, Belgium) equilibrated with Binding Buffer A.

Article Title: Overlapping Specificity of Duplicated Human Pancreatic Elastase 3 Isoforms and Archetypal Porcine Elastase 1 Provides Clues to Evolution of Digestive Enzymes *
Article Snippet: Based on the sequence pattern of the inhibitor binding loops selected against the human elastases, 12 different inhibitor variants were designed and created using PCR-based mutagenesis ( ). .. The recombinant SGPI-2 variants were expressed into the cytoplasm of E. coli SHuffle T7 cells (C3026, New England Biolabs) fused to the C terminus of the E. coli protein-disulfide isomerase DsbC following the strategy of Nozach et al. ( ).

Mutagenesis:

Article Title: Overlapping Specificity of Duplicated Human Pancreatic Elastase 3 Isoforms and Archetypal Porcine Elastase 1 Provides Clues to Evolution of Digestive Enzymes *
Article Snippet: Based on the sequence pattern of the inhibitor binding loops selected against the human elastases, 12 different inhibitor variants were designed and created using PCR-based mutagenesis ( ). .. The recombinant SGPI-2 variants were expressed into the cytoplasm of E. coli SHuffle T7 cells (C3026, New England Biolabs) fused to the C terminus of the E. coli protein-disulfide isomerase DsbC following the strategy of Nozach et al. ( ).

Subcloning:

Purification:

Article Title: Overlapping Specificity of Duplicated Human Pancreatic Elastase 3 Isoforms and Archetypal Porcine Elastase 1 Provides Clues to Evolution of Digestive Enzymes *
Article Snippet: Paragraph title: Expression and Purification of SGPI-2 Variants ... The recombinant SGPI-2 variants were expressed into the cytoplasm of E. coli SHuffle T7 cells (C3026, New England Biolabs) fused to the C terminus of the E. coli protein-disulfide isomerase DsbC following the strategy of Nozach et al. ( ).

Article Title: Plasmodium falciparum MSP3 Exists in a Complex on the Merozoite Surface and Generates Antibody Response during Natural Infection
Article Snippet: Paragraph title: Expression and purification of recombinant proteins. ... The large N-terminal fragment of MSP3N in a pET-28a expression construct was transformed into SHuffle T7 competent E. coli cells (NEB) for expression; the cells were induced with 0.5 mM isopropyl- d -thiogalactopyranoside (IPTG) (Sigma Chemical Co.) for 8 h at 30°C.

Article Title: Nuclear Import of Arabidopsis Poly(ADP-Ribose) Polymerase 2 Is Mediated by Importin-α and a Nuclear Localization Sequence Located Between the Predicted SAP Domains
Article Snippet: Paragraph title: Protein Expression and Purification ... For protein expression in E. coli , pOPINS3C carrying His6:SUMO:SAP and pOPINE carrying the α-GFP-nanobody:Halo:His6 construct (Addgene plasmid #111090) were transformed into SHuffle® T7 Competent E. coli cells (New England Biolabs).

Sequencing:

Article Title: Overlapping Specificity of Duplicated Human Pancreatic Elastase 3 Isoforms and Archetypal Porcine Elastase 1 Provides Clues to Evolution of Digestive Enzymes *
Article Snippet: Based on the sequence pattern of the inhibitor binding loops selected against the human elastases, 12 different inhibitor variants were designed and created using PCR-based mutagenesis ( ). .. The recombinant SGPI-2 variants were expressed into the cytoplasm of E. coli SHuffle T7 cells (C3026, New England Biolabs) fused to the C terminus of the E. coli protein-disulfide isomerase DsbC following the strategy of Nozach et al. ( ).

Article Title: The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall
Article Snippet: The fragment was ligated into pET28a(+) expression vector (Novagen), creating a sequence with an N-terminal histidine tag. .. Cloning was done using E. coli DH5α and selection with 30 µg/ml kanamycin; the final expression vector was transformed into SHuffle T7 competent E. coli cells (New England Biolabs).

Selection:

Lysis:

Article Title: Bifunctional Chloroplastic DJ-1B from Arabidopsis thaliana is an Oxidation-Robust Holdase and a Glyoxalase Sensitive to H2O2
Article Snippet: This expression vector was transformed into SHuffle® T7 competent E. coli (New England Biolabs, Ipswitch, MA, USA) and plated on agar to obtain a single colony, which was grown overnight at 30 °C in Luria-Bertani Broth (LB) supplemented with 100 μg/mL ampicillin. .. Cells were pelleted and resuspended in Lysis Buffer, lysed in a cell cracker at 20 kilopounds per square inch, and centrifuged at 40,000 × g for 30 min, 4 °C.

Article Title: Plasmodium falciparum MSP3 Exists in a Complex on the Merozoite Surface and Generates Antibody Response during Natural Infection
Article Snippet: The large N-terminal fragment of MSP3N in a pET-28a expression construct was transformed into SHuffle T7 competent E. coli cells (NEB) for expression; the cells were induced with 0.5 mM isopropyl- d -thiogalactopyranoside (IPTG) (Sigma Chemical Co.) for 8 h at 30°C. .. The induced cell pellet was disrupted by sonication in Tris-NaCl lysis buffer (0.05 M Tris, pH 8.0, and 0.15 M NaCl, 0.01 M dithiothreitol [DTT], 1 mM phenylmethylsulfonyl fluoride [PMSF], 1% Triton X-100) with 9-s pulses at 9-s intervals 10 times using a miniprobe.

Article Title: Bifunctional Chloroplastic DJ-1B from Arabidopsis thaliana is an Oxidation-Robust Holdase and a Glyoxalase Sensitive to H2O2
Article Snippet: This expression vector was transformed into SHuffle® T7 competent E. coli (New England Biolabs, Ipswitch, MA, USA) and plated on agar to obtain a single colony, which was grown overnight at 30 °C in Luria-Bertani Broth (LB) supplemented with 100 μg/mL ampicillin. .. Cells were pelleted and resuspended in Lysis Buffer, lysed in a cell cracker at 20 kilopounds per square inch, and centrifuged at 40,000 × g for 30 min, 4 °C.

SDS Page:

Plasmid Preparation:

Article Title: Efficient biosynthesis of heterodimeric C3-aryl pyrroloindoline alkaloids
Article Snippet: Finally, we cloned trx-fd and mbp-fdr into a pRSFduet vector and nascB into pET21a to achieve the co-expression of Fd, FdR, and NascB in E. coli . .. The B type strains are often used for protein expression, while K type strains are mostly used for DNA cloning but also for protein expression, such as Shuffle T7 competent E. coli from New England Biolabs.

Article Title: Nuclear Import of Arabidopsis Poly(ADP-Ribose) Polymerase 2 Is Mediated by Importin-α and a Nuclear Localization Sequence Located Between the Predicted SAP Domains
Article Snippet: .. For protein expression in E. coli , pOPINS3C carrying His6:SUMO:SAP and pOPINE carrying the α-GFP-nanobody:Halo:His6 construct (Addgene plasmid #111090) were transformed into SHuffle® T7 Competent E. coli cells (New England Biolabs). .. The His6:ΔIBB-IMPORTIN-α2 protein was expressed from pOPINF in SoluBL21 cells (Genlantis).

Positron Emission Tomography:

Produced:

Article Title: The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall
Article Snippet: Recombinant Gas1p was produced using a Pichia pastoris expression system ( ). .. Cloning was done using E. coli DH5α and selection with 30 µg/ml kanamycin; the final expression vector was transformed into SHuffle T7 competent E. coli cells (New England Biolabs).

Concentration Assay:

Article Title: The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall
Article Snippet: Cloning was done using E. coli DH5α and selection with 30 µg/ml kanamycin; the final expression vector was transformed into SHuffle T7 competent E. coli cells (New England Biolabs). .. At an optical density of 0.8, the culture was induced with a final concentration of 1 mM isopropyl-β-d -1-thiogalactopyranoside (Sigma-Aldrich).

Recombinant:

Article Title: Overlapping Specificity of Duplicated Human Pancreatic Elastase 3 Isoforms and Archetypal Porcine Elastase 1 Provides Clues to Evolution of Digestive Enzymes *
Article Snippet: .. The recombinant SGPI-2 variants were expressed into the cytoplasm of E. coli SHuffle T7 cells (C3026, New England Biolabs) fused to the C terminus of the E. coli protein-disulfide isomerase DsbC following the strategy of Nozach et al. ( ). .. The construct also incorporated an N-terminal His tag and a TEV proteinase cleavage site between the inhibitor and DsbC.

Article Title: The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall
Article Snippet: Paragraph title: Production of recombinant Gas1p and Bgl2p. ... Cloning was done using E. coli DH5α and selection with 30 µg/ml kanamycin; the final expression vector was transformed into SHuffle T7 competent E. coli cells (New England Biolabs).