escherichia coli shuffle t7 express lysy  (New England Biolabs)

Journal: Biotechnology Letters

Article Title: Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli

doi: 10.1007/s10529-013-1180-z

Figure Lengend Snippet: Cytoplamic expression of proTHI-TRX fusion proteins in strain C3030. For all fusion proteins, 1 ml of cell culture was pelleted and dissolved in 100 μl sample buffer. 10 μl from this extract and an equivalent amount for total soluble cytoplasmic fractions were separated on Tricine/SDS gels. (M) Protein marker, ( 1 ) un-induced crude extract, ( 2 ) induced crude extract, ( 3 ) total soluble fraction taken after cell lysis by sonication, ( 4 ) insoluble fraction after sonication. Red stars indicate the 25 kDa fusion protein

Article Snippet: The amount of fusion protein that could be produced from the SHuffle strain C3030 is shown in Table and was higher than obtained with Rosetta(DE3)pLysS.

Techniques: Expressing, Cell Culture, Marker, Lysis, Sonication

Journal: Biotechnology Letters

Article Title: Comparison of periplasmic and intracellular expression of Arabidopsis thionin proproteins in E. coli

doi: 10.1007/s10529-013-1180-z

Figure Lengend Snippet: Comparison of Ni–NTA purified proTHI-TRX fusion proteins from strain C3030. a Coomassie Brilliant Blue staining. b Western blot with anti-His tag antibody. Each well contained 3 μg protein. (M) Prestained protein marker, ( 1 ) proTHI2.1-TRX, ( 2 ) proTHI2.2-TRX, ( 3 ) proTHI2.3-TRX, ( 4 ) proTHI2.4-TRX

Article Snippet: The amount of fusion protein that could be produced from the SHuffle strain C3030 is shown in Table and was higher than obtained with Rosetta(DE3)pLysS.

Techniques: Purification, Staining, Western Blot, Marker

Journal: Infection and Immunity

Article Title: Evaluation of a Plasmodium-Specific Carrier Protein To Enhance Production of Recombinant Pfs25, a Leading Transmission-Blocking Vaccine Candidate

doi: 10.1128/IAI.00486-17

Figure Lengend Snippet: Expression of chimeric r Pf s25/8(CΔS) and unfused r Pf s25. (A) Schematic depiction of expression constructs for the production of r Pf s25/8(CΔS) and r Pf s25. Expression plasmids for chimeric r Pf s25/8(CΔS) or unfused r Pf s25 were transformed into E. coli SHuffle T7 Express lysY cells. (B and C) r Pf s25/8(CΔS) lysates harvested before ( T 0 ) or 3 h after ( T 3 ) induction were separated by SDS-PAGE (10% gel) under reducing conditions, followed by Coomassie blue staining (B) or immunoblot analysis (C) using rabbit-anti-r Pf MSP8 IgG. The asterisk highlights r Pf s25/8(CΔS) at the predicted size. (D and E) The expression of unfused r Pf s25 was assessed as described above, on 12% polyacrylamide gels under reducing conditions, followed by Coomassie blue staining (D) or immunoblot analysis (E) using an anti-His MAb. The asterisk highlights r Pf s25 at the predicted size.

Article Snippet: The r Pf s25 expression construct was transformed into SHuffle T7 Express lysY competent E. coli cells (New England BioLabs).

Techniques: Expressing, Construct, Transformation Assay, SDS Page, Staining

Journal: Microbial Cell Factories

Article Title: A high-throughput expression screening platform to optimize the production of antimicrobial peptides

doi: 10.1186/s12934-017-0637-5

Figure Lengend Snippet: Expression screening in E. coli . Expression screening was carried out in 96-well microtiter plates to identify high producers for three different target proteins. The promoter, protein fusion partner and E. coli strain were varied. a IMPI, b BR021, c AFP. Rosetta-gami 2 Rosetta-gami 2 (DE3) pLysS, Origami Origami 2, SHuffle SHuffle T7 Express lysY, BL21 BL21 (DE3), C41 OverExpress C41 (DE3) pLysS, C43 OverExpress C43 (DE3) pLysS

Article Snippet: Transformation of E. coli expression strains Six different E. coli strains were used for expression screening: Rosetta-gami 2 (DE3) pLysS (Merck Millipore), Origami 2 (Merck Millipore), BL21 (DE3) (NEB), SHuffle T7 Express lysY (NEB), OverExpress C41 (DE3) pLysS (Sigma-Aldrich), OverExpress C43 (DE3) pLysS (Sigma-Aldrich).

Techniques: Expressing

Journal: Frontiers in Plant Science

Article Title: Molecular and Biochemical Analysis of Duplicated Cytosolic CuZn Superoxide Dismutases of Rice and in silico Analysis in Plants

doi: 10.3389/fpls.2022.864330

Figure Lengend Snippet: Analysis of oxidative stress tolerance of E. coli SHuffle T7 Express cells overexpressing OsCSD1 and OsCSD4. (A) Comparative analysis of growth of E. coli cells (absorbance at 600 nm) containing pET28a (empty vector), pET28a-OsCSD1, and pET28a-OsCSD4 plasmids. Cells induced with IPTG were treated with increasing methyl viologen concentration (0–0.500 mM) and monitored spectrophotometrically. The experiment was repeated three times and data are represented as mean value ± SD. Statistical significance is indicated by * p

Article Snippet: Briefly, E. coli SHuffle T7 Express cells containing recombinant pET28a-OsCSD1 or pE28a-OsCSD4 plasmid were grown overnight at 30°C in LB medium containing kanamycin (25 μg ml–1 , LB-Kan+ ).

Techniques: Plasmid Preparation, Concentration Assay