Isolation And Characterization Of Transmissible Gastroenteritis Virus Of Swine And Its Gene 7 Cloning And Construction Of Prokaryotic Expression Vector

Transmissible Gastroenteritis is a kind of highly contagious disease. The clinical manifestations of typical TGE is serious diarrhea, sickness and dehydration. Transmissible Gastroenteritis Virus of Swine(TGEV) belongs to the genus Coronavirus, family coronaviridae. The virus causes transmissible gastroenteritis in all kinds and ages of pigs. Particularly, its mortality in pigs under 2 weeks old may reach 100%.In this research, Transmissible Gastroenteritis Virus was successfully isolated from diarrheal piglet fecal sample of a farm in Sichuan Province. The virus was replication in ST cells and resulted in cytopathogenic effect(CPE) after two generations. Then the Transmissible Gastroenteritis Virus (TGEV) strain was named TGEV SC-1,which was isolated successfully after observation and detection under the lectron microscope and the virus was about 90 nm in diameter;organic reagent sensitivity experiment by chloroform (sensitiveness);the nucleic acid type experiment(RNA);TCID50 detection(10^-4.61/50 μ l);virus neutralization experiment (index number is 128. 8)and so on.According to these publiced gene 7 sequence in the GenBank, one pair primers were designed bythen succeed in the clone of 303bp long fragment(GenBank accession number is DQ437507). Alignment of pair of sequence and analysis of the putative amino acid sequence indicates that there are 92.4% a similary with other Trassimissible gastroenteritis virus strains. And last, we are successful in cloning the fragment into Prokaryotic Expression Vector PBV220.