Li Yan, Nie Mingfei, Wei Wei, Wen Kai, Jia Ying, Huo Hui, Wang Junwei
College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
Sheng Wu Gong Cheng Xue Bao. 2010 Mar;26(3):311-6.
In this study, we cloned the NS3 gene from bovine viral diarrhea virus (BVDV) VEDEVAC strain. The result showed that the average P-distance of Pestivirus NS3 amino acid sequence was 0.07 and the VEDEVAC strain was classified to BVDV type 1. Using pET-30a(+) as vector and Escherichia coli Rosetta (DE3) as host, we obtained purified recombinant NS3 protein by Ni-NTA affinity chromatography. Western blotting analysis demonstrated that both BVDV positive serum and classical swine fever virus (CSFV) positive serum were able to recognize the recombinant NS3 protein. Indirect-ELISA assay indicated that the protein could be used as detection antigen.