[Sequence analysis, expression and antigenicity detection of bovine viral diarrhea virus NS3 gene].
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.
Li
Yan
Y
College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
Nie
Mingfei
M
Wei
Wei
W
Wen
Kai
K
Jia
Ying
Y
Huo
Hui
H
Wang
Junwei
J
D000818:Animals; D002417:Cattle; D003001:Cloning, Molecular; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics* / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D010447:Peptide Hydrolases / Q000235:genetics* / Q000276:immunology*; D010802:Phylogeny; D020365:RNA Helicases / Q000235:genetics* / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D020539:Sequence Analysis, Protein; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000276:immunology*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D010447:Peptide Hydrolases; D020365:RNA Helicases; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; C086730:p80 protein, bovine viral diarrhea virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases