关键词: MuIFN-γ, EMCVVP1, 原核表达, 免疫佐剂

Abstract: Objective Study the molecular adjuvant effect of MuIFN-T by co-expressing MuIFN-T and EMCV structural protein VP1. Method Mature peptide gene of MuIFN-T was cloned by using RT-PCR. VP1 of pig EMCV was subcloned from pGEX-6p-l/vpl, and constructed into the express plasmid pET32-a /VPl/Rosetta(DE3) and co-expression plasmid pET32-a/VPl/MuIFN-T, respectively. BALB/c mice were inoculated with the VP1 and VIuIFN-Y/VPl proteins in 'E.coli, respectively, for three times at two week intervals, serum IgG were detected by ELISA every two weeks post-inoculation. Result In Ecoli Rosetta(DE3), expression and co-expression were succeeded IPTG (Isopropyl β-D-l-Thiogalactopyranoside) induction, with 60 MW of VP1 protein and 73 MW of co-expressed protein. The serum IgG of mice post-inoculated by VP1 and MuIFN-T/VP 1 are 1 : 32000 and 1 : 128000. Conclusions VP1 had good antigenicity which could stimulated humoral immune response in inoculated mice, the detected serum IgG of MuIFN-γ/VP 1 inoculation is higher than VP1. Moreover, immune enhancing effects was found in mice inoculated with co-expressed MuIFN-T/VPl .LMuIFN-T was a satisfactory immune adjuvant.

Key words: MuIFN-γ, EMCV VP1, Prokaryotic expression, Immune adjuvant