[Infectious bovine rhinotracheitis viral gG expression and gG-ELISA development].
Taking the genome DNA of Infectious Bovine Rhinotracheitis Virus (IBRV) as the template, the gG gene was amplified with PCR and cloned into the T cloning vector pMD18-T. After being identified by restriction digestion and DNA sequencing, the insert was subcloned into the expression vector pGEX-KG. Sodium docecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot assay showed that this gene was expressed as both soluble form and inclusion body by the transformed E. coli BL21 strain (DE3). The fusion protein was purified and used as the coating antigen to develop the indirect Enzyme-Linked Immunosorbent Assay (ELISA). Comparison between this gG-ELISA and commercial IBRV gB-ELISA Kit (IDEXX) was made in the detection of 380 cow serum samples. The results demonstrated an agreement of 92%. By using this novel gG-ELISA, 1248 cow serum samples were tested and the average positive rate of IBRV antibodies for imported cows is 21.7%, while the positive rate ranged greatly from 0.0%-41.5% for Hubei local Chinese Black and White Dairy Cows.
The State Key Laboratory of Agricultural Microbiology, Wuhan 430070, China.
Bang-Fen
BF
Yan
Zeng
Z
Chen
Shu-Huan
SH
Zhang
Xiang-Mei
XM
Lin
Ying-Yu
YY
Chen
Yan-Jie
YJ
Chao
De-Xue
DX
Li
Nian-Hua
NH
Song
Huan-Chun
HC
Chen
Ai-Zhen
AZ
Guo
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D002417:Cattle; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D007242:Herpesvirus 1, Bovine / Q000235:genetics* / Q000276:immunology; D008297:Male; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins