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重组杆状病毒表达的伪狂犬病病毒糖蛋白gII的特性分析

Characterization of pseudorabies virus glycoprotein gII expressed by recombinant baculovirus.

作者信息

Xuan X, Nakamura T, Ihara T, Sato I, Tuchiya K, Nosetto E, Ishihama A, Ueda S

机构信息

Nippon Institute for Biological Science, Tokyo, Japan.

出版信息

Virus Res. 1995 May;36(2-3):151-61. doi: 10.1016/0168-1702(94)00112-p.

Abstract

The gene encoding the complete glycoprotein gII (homologue of gB of herpes simplex virus) of pseudorabies virus (PrV) was inserted into a baculovirus transfer vector, and a recombinant virus expressing gII was isolated. Three gII-related recombinant baculovirus-expressed peptides of 100, 60, and 45 to 50 kDa were detected with a polyclonal antibody against gII; these correspond to the authentic subunits gIIa and its cleavage products gIIb and gIIc, respectively. These proteins were subjected to N-terminal sequencing, and the results showed that the protease cleavage sites were identical to those of authentic gII. The expressed gII was shown to be transported to the surface of infected cells as judged by an indirect immunofluorescence test. Antibodies raised in mice immunized with the recombinant gII neutralized the infection of PrV in vitro. Mice inoculated with the recombinant gII were completely protected from lethal challenge with PrV.

摘要

将编码伪狂犬病病毒(PrV)完整糖蛋白gII(单纯疱疹病毒gB的同源物)的基因插入杆状病毒转移载体中,分离出表达gII的重组病毒。用抗gII的多克隆抗体检测到三种与gII相关的杆状病毒表达的重组肽,大小分别为100、60和45至50 kDa;它们分别对应于天然亚基gIIa及其切割产物gIIb和gIIc。对这些蛋白质进行了N端测序,结果表明蛋白酶切割位点与天然gII的相同。通过间接免疫荧光试验判断,所表达的gII被转运至感染细胞的表面。用重组gII免疫的小鼠产生的抗体在体外中和了PrV的感染。接种重组gII的小鼠完全受到保护,免受PrV致死性攻击。

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