هم سانه سازي و بيان ژن زيرواحدB توكسين حساس به حرارت (LTB) باكتري اشريشياكلي انتروتوكسيژنيك به عنوان كانديداي واكسن

Cloning and expression of enterotoxigenic Escherichia coli heat labile toxin B subunit (LTB) as a vaccine candidate

Aims. Enterotoxigenic Escherichia coli (ETEC) is the most common agent of diarrhea disease among other bacterial agents and mortality rate of this disease is high in worldwide. Designing and producing vaccine against this disease is one of the purposes of world health organization. Heat-labile toxin (LT) is one of the most important virulence agents of bacteria and a vaccine candidate. The aim of this study was the expression of heat labile B subunit toxin (LTB) to investigate its immunologicl property. Materials & Methods. Information about LTB gene was obtained from gene bank and appropriate primers were designed accordingly. Genomic PCR reaction was performed and its product was cloned into pBlueskriptll SK cloning vector and pET28a expression vector using restriction enzymes and was transformed to competent cells. Then, LTB gene was expressed by induction of IPTG. The concentration of protein was determined and confirmed by immunobloting after SDS-PAGE. Results. 15.5 kD protein (MWt) was observed on SDS-PAGE comparing to noninduced sample and was confirmed with anti-cholera toxin B subunit antibody. Conclusion. Expressed LTB protein could be used in vaccine designing and also as strong mucosal immune system adjuvant.