Production of Bovicin HC۵ in two strains of E. coli, Rosetta gami and Rosetta plysS

Background and Aim : Introduction: Bacteriocins are antimicrobial peptides that are produced by ribosomes in bacteria and some archaea. Evidence suggests that these peptides could be used to treat cancer disease. Bovicin HC۵ is a bacteriocin produced by Streptococcus bovis HC۵ and was used in the treatment of liver cancer. Objective: The expression of recombinant bovicin HC۵ was performed for the first time in two bacterial strains of E. coli, Rosetta gami, and Rosetta plysS. Methods : The cDNA of Bovicin HC۵ was cloned in the pET۲۱a vector. For protein expression, IPTG with ۰.۵ and ۱ mM concentrations was used. Also, cultivation temperature of ۳۵ °C and post-induction times of ۲, ۴, and ۸ hours were selected for incubation. For protein expression analysis, the dot blot method was used and then the data were quantified by ImageJ software.Results : : The highest expression level, ۲۰.۴۹%, in the Rosetta gami strain was achieved at ۰.۵ mM IPTG and a post-induction time of ۴ hours. Also, the maximum expression rate, ۲۱.۵۰%, was observed in Rosetta plysS at ۰.۵ mM IPTG and a post-induction time of ۸ hours.Conclusion : According to the results, both strains had appropriate protein expression levels.

کلیدواژه ها:

bovicin HC۵ ، E. coli ، protein expression