Nucleotide sequence analysis and expression of bovid herpesvirus 1 glycoproteins

通过在病毒接种中加入肝素和用BHV-L的主要糖蛋白的肝素酶处理细胞，可以阻止牛疱疹病毒I（by -1）吸附和感染细胞，只有gIII被发现特异性地与肝素结合。针对抗原位点la的声克隆抗体抑制了gIII的结合，干扰了病毒的吸附。这些发现表明，病毒对细胞的吸附是通过gIII抗原位点la与细胞表面的肝素样片段相互作用介导的，肝素样片段是BUY-1的受体。将gIII基因克隆到表达载体pcDLSRalpha-456质粒中，利用得到的质粒pSRBR3转染COS-7细胞。用超声克隆抗体对转染的细胞进行荧光抗体染色，证实了抗原的真实性，并且发现糖蛋白- C57BL小鼠红细胞表面表达在表达gIII的细胞上。阻断病毒吸附的单克隆抗体和肝素可特异性抑制其吸血活性。这些发现表明，glll与红细胞结合，并可能与宿主细胞结合，而不与其他病毒成分结合。使用4- base识别限制性内切酶，我们试图比较BRY-1分离株的基因组结构，尽管糖蛋白的核苷酸测序工作尚未完成。结果表明，4碱基识别核酸内切酶指纹图谱不仅可以作为鉴定各BHY-L菌株的有效工具，而且可以推测各菌株之间的流行病学关系。

Bovid herpesvirus I(BBY-1)was prevented from adsorbing to and infecting cells by addition of heparin to the virus inoculus and by treatment of the cells with heparinase- of the major glycoproteins of BHV-L only gIII was found to bind specifically to heparin. The binding of gIII was inhibited by a sonoclonal antibody against antigenic site la, which interferes with the adsorption of the virus. These findings indicate that the virus adsorption to cells is mediated by interaction of the gIII antigenic site la with a heparin-like moiety on the cell surface, which serves as a receptor for BUY-1. The gIII gene was cloned into expression vector plassid pcDLSRalpha-456 and a resulting plassid pSRBR3 were used, -to transfect COS-7 cells. Fluoresent antibody staining with sonoclonal antibodies on the transfected cells demonstrated antigenic authenticity as well as surface expression of the glycoprotein- C57BL mouse crythrocytes were found to adsorb onto the gIII -expressing cells. The hemaasorbing activity was specifically inhibited by the monoclonal antibody blocking virus adsorption and by heparin. Theses findings indicate that the glll bind to the erythr ocytes and probably to the host cells in no conjunction with other viral components.Using 4-bese-recognition restricted endonuclease we attempted to compare genome construction among BRY-1 isolates, although nucleotide sequencing work on the glycoproteins was not done. Results suggested that the 4-basc recognition endonuclease fingerprint could be a useful tool not only discriminate each BHY-L strain also to presume epidemiological relationship among each isolate.