Enhanced expression of recombinant beta toxin of <i>Clostridium perfringens</i> type B using a commercially available <i>Escherichia coli</i> 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 RosettaTM(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 RosettaTM(DE3). Recombinant protein was expressed as a soluble protein after isopropyl β-D-1-thiogalactopyranoside (IPTG) induction in strain RosettaTM(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.

Recombinant Alpha, Beta, and Epsilon Toxins of Clostridium perfringens: Production Strategies and Applications as Veterinary Vaccines

“…Further studies have found that the strain of the E. coli can influence the solubility of the protein. For example, Bakhshi et al [74] found that the rCPB expressed in BL21 (DE3) was insoluble, while that expressed in Rosetta (DE3) was soluble. Reducing the expression temperature (to between 16 and 28 °C) is a common approach by which researchers try to increase protein solubility.…”

Vaccination against pathogenic clostridia in animals: a review

“…In recent years, research has been focused on experimental recombinant C. perfringens vaccines. The ETX and cpb gene regions of the C. perfringens have been used for design of the new generation vaccines, separately (Aziminia et al 2016 ; Bakhshi et al 2016 ). Recombinant vaccines expressed in E. coli have resulted in high immunity in laboratory animals (Aziminia et al 2016 ; Lobato et al 2010 ).…”

Molecular Cloning of a New Bi-Functional Fusion Protein of Clostridium perfringens Type A Alpha and Clostridium septicum Alpha Toxin Genes in E. coli

“…After amplification by PCR reaction, all amplicons were electrophoresed for their qualification in gel electrophoresis (1.0% agarose (Sinaclon, Iran)) (20). The resulting PCR products (about 1.2 kb) were purified by Gel Extraction Kit (Vivantis, Malaysia) according to the manufacturer's recommendations (21,22).…”