Prokaryotic Expression of Murine Cellular Prion Protein for in Vitro Evaluation

Abstract

M cell (Microfold cell) targeting is a potential strategy for oral vaccine development due to its role in presenting antigen to mucosal lymphoid tissue. Numerous researches have focused on directing vaccine antigens to M cell receptors, based on their affinity with specific ligands. This study was aimed to biosynthesize murine cellular prion protein (mPrP C ), a highly expressed receptor on the M cell apical surface. The mPrP C coding sequence was obtained by PCR for construction of pGEX-5X-1-mprpc plasmid. Then, recombinant plasmid was induced for expression in E. coli BL21(DE3) cells by IPTG. The overexpression of mPrP C was confirmed by SDS-PAGE and Western blotting. Since the expressed protein was found to remain mostly in insoluble fraction, DTT and glycerol were used to improve the solubility. This soluble, recombinant mPrP C could be used for in vitro evaluation, and also on further research for developing antigen targeting M cell.