Research Note

Enhanced expression of recombinant beta toxin of Clostridium perfringens type B using a commercially available Escherichia coli strain

Fatemah Bakhshi, Reza Pilehchian Langroudi, Bahram Golestani Imani
Onderstepoort Journal of Veterinary Research | Vol 83, No 1 | a1136 | DOI: https://doi.org/10.4102/ojvr.v83i1.1136 | © 2016 Fatemah Bakhshi, Reza Pilehchian Langroudi, Bahram Golestani Imani | This work is licensed under CC Attribution 4.0
Submitted: 18 December 2015 | Published: 30 June 2016

About the author(s)

Fatemah Bakhshi, Department of Biology, Islamic Azad University, Urmia branch, Iran, Islamic Republic of
Reza Pilehchian Langroudi, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Alborz, Karaj, Iran, Islamic Republic of
Bahram Golestani Imani, Department of Biology, Islamic Azad University, Urmia branch, Urmia, Iran, Islamic Republic of

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.

Keywords: C. perfringens beta toxin (CPB); expression; RosettaTM; BL21


Keywords

C. perfringens beta toxin (CPB); expression; RosettaTM; BL21

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