Eukaryotic Expression Of S Gene Of A,D Sites Of TGEV

Procine transmissible gastroenteritis virus(TGEV) causes transmissible gastroenteritis(TGE) in all kinds and ages of pigs. Particularly, its mortality in pigs under 2 week old may reach 100%. TGE is a kind of acute highly infectious disease . The clinical manifestations of typical TGE consist of omitting, serious diarrhea and loss of water. It is an important disease for the development of pigs. Accurate diagnosis and effective vaccination are main means for TGE . S protein is the only structure protein to induce neutral antibody. So the expression of S gene is important for TGE. In this study, One pair of primers are designed according to the published sequence of TH-98 S gene with primer 5.0 software. The product of PCR is including sites A and D of S gene. Which is named S1. Then the S1 gene is cloned into the vector pMD18-T. Identification of restriction enzyme, PCR and sequencing indicate that the S1 gene has been cloned successfully, S1 gene is inserted into BamHâ… and Hindâ…¢multiple cloning sites of the vector pMel-Bac C, which is named pMel-Bac C-S1, Bac-N-Blue DNA and pMel-Bac C-S1 are contransinfected into insect cell Sf9, By plaque assay, the recombinant baculovirus are obtained, The techniques of SDS-PAGE and Western-blot are used to detect and identify the products expressed in Sf9 lines. The MW of the recombinants protein is 37KDa. The mount of protein evaluated by densitometric scanning can be reached 8.3%. The indirect enzyme linked immunosorbent assay(ELISA) shows that the recombinant protein not only has reactivity to react with antibody, but also prossesse strong immunogenicity to induce production of S antibody. It is the first time reported that S gene of TGEV is cloned and expressed in Baculovirus Expression System. The study provides the important basis for further reseach of TGEV molecular peculiarity.