使用针对细菌病毒融合蛋白的抗血清检测小鼠肝炎病毒非结构蛋白。

Detection of mouse hepatitis virus nonstructural proteins using antisera directed against bacterial viral fusion proteins.

作者信息

Zoltick P W, Leibowitz J L, DeVries J, Pachuk C J, Weiss S R

机构信息

Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia 19104-6076.

出版信息

Adv Exp Med Biol. 1990;276:291-9. doi: 10.1007/978-1-4684-5823-7_40.

DOI:10.1007/978-1-4684-5823-7_40

PMID:1966415

Abstract

Mouse hepatitis virus, strain A59 cDNAs were inserted into the procaryotic fusion vector pGE374. RecA/viral/LacZ tripartite fusion proteins were synthesized from these plasmids and purified from E. coli. Antisera were raised in rabbits against these fusion proteins. Viral nonstructural proteins were detected in infected murine fibroblasts and glial cells. The anti-gene B, ORF1 sera detect a 30K cytoplasmic protein while the anti-gene E, ORF2 sera detect a 9.6K protein. Sera raised against proteins encoded in cDNAs from 5' portions of gene A immunoprecipitate the 200-250K polypeptides synthesized in vitro from genome RNA. Antisera raised against proteins encoded in both 5' and 3' portions of gene A immunoprecipitate membrane associated polypeptides of 150K and greater than 600K from MHV infected cells.

摘要

将小鼠肝炎病毒A59株的cDNA插入原核融合载体pGE374。从这些质粒中合成RecA/病毒/LacZ三方融合蛋白，并从大肠杆菌中纯化。用这些融合蛋白对兔子进行免疫以产生抗血清。在感染的小鼠成纤维细胞和神经胶质细胞中检测到病毒非结构蛋白。抗基因B、ORF1血清检测到一种30K的细胞质蛋白，而抗基因E、ORF2血清检测到一种9.6K的蛋白。针对基因A 5'部分cDNA编码的蛋白产生的血清可免疫沉淀从基因组RNA体外合成的200 - 250K多肽。针对基因A 5'和3'部分编码的蛋白产生的抗血清可从感染MHV的细胞中免疫沉淀150K和大于600K的膜相关多肽。