title: Enhanced expression of recombinant beta toxin of Clostridium perfringens type B using a commercially available Escherichia coli strain.
abstract: Clostridium perfringens beta toxin is only produced by types B and C and plays an important role in many human and animal diseases, causing fatal conditions that originate in the intestines. We compared the expression of C. perfringens type B vaccine strain recombinant beta toxin gene in the Escherichia coli strains Rosetta(DE3) and BL21(DE3). The beta toxin gene was extracted from pJETβ and ligated with pET22b(+). pET22β was transformed into E. coli strains BL21(DE3) and Rosetta(DE3). Recombinant protein was expressed as a soluble protein after isopropyl β-D-1-thiogalactopyranoside (IPTG) induction in strain Rosetta(DE3) but not in BL21(DE3). Expression was optimised by growing recombinant cells at 37 °C and at an induction of 0.5 mM, 1 mM, 1.5 mM IPTG. Expression was evaluated using sodium dodecyl sulfate Polyacrylamide gel electrophoresis (SDS-PAGE). The recombinant protein was purified via Ni-NTA and was analysed using western blot. We concluded that E. coli strain RosettaTM(DE3) can enhance the expression of C. perfringens recombinant beta toxin.
authors.forename: Fatemah
authors.initials: F
authors.lastname: Bakhshi
authors.affiliation: Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Alborz, Karaj. rpl4321@gmail.com.
authors.forename: Reza
authors.initials: R
authors.lastname: Pilehchian Langroudi
authors.forename: Bahram Golestani
authors.initials: BG
authors.lastname: Eimani
mesh_terms: D001427:Bacterial Toxins / Q000235:genetics* / Q000378:metabolism; D001428:Bacterial Vaccines / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western / Q000662:veterinary; D003016:Clostridium perfringens / Q000235:genetics* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA / Q000662:veterinary
chemical_list: D001427:Bacterial Toxins; D001428:Bacterial Vaccines; C031884:CPB protein, Clostridium perfringens; D011994:Recombinant Proteins
keywords: C. perfringens beta toxin (CPB); expression; RosettaTM; BL21.