传染性胃肠炎病毒强毒株的整合膜蛋白：分子特征、序列及在大肠杆菌中的表达
The integral membrane protein from a virulent isolate of transmissible gastroenteritis virus: molecular characterization, sequence and expression in Escherichia coli.
作者信息
Britton P, Cármenes R S, Page K W, Garwes D J
机构信息
Division of Microbiology, AFRC Institute for Animal Health, Compton Laboratory, Berkshire, UK.
出版信息
Mol Microbiol. 1988 Jul;2(4):497-505. doi: 10.1111/j.1365-2958.1988.tb00056.x.
Abstract
Subgenomic mRNA from a virulent isolate of porcine transmissible gastroenteritis virus (TGEV) was used to produce cDNA clones. Part of a new clone and a previously reported clone were sequenced and used to construct the viral gene for integral membrane protein. A single open reading frame (ORF) encoding a polypeptide of 262 amino acids, relative molecular mass (Mr) 29,459, was identified. The positive identification of the polypeptide as the integral membrane protein was demonstrated by the production in E. coli of a chimaeric protein comprising most of the ORF encoding the Mr 29,459 polypeptide and beta-galactosidase. The chimaeric protein reacted with a specific monoclonal antibody to viral integral membrane protein and antibodies raised against the chimaeric protein immune precipitated the viral protein. Comparison with the sequence of an avirulent isolate indicates amino acid residues that may be important in pathogenicity.
摘要
来自猪传染性胃肠炎病毒（TGEV）强毒株的亚基因组mRNA被用于制备cDNA克隆。对一个新克隆的部分序列和先前报道的一个克隆进行了测序，并用于构建完整膜蛋白的病毒基因。鉴定出一个单一的开放阅读框（ORF），其编码一个由262个氨基酸组成、相对分子质量（Mr）为29459的多肽。通过在大肠杆菌中产生一种包含编码Mr 29459多肽的大部分ORF和β-半乳糖苷酶的嵌合蛋白，证实了该多肽作为完整膜蛋白的阳性鉴定。该嵌合蛋白与针对病毒完整膜蛋白的特异性单克隆抗体发生反应，并且针对该嵌合蛋白产生的抗体免疫沉淀了病毒蛋白。与无毒株序列的比较表明了可能在致病性中起重要作用的氨基酸残基。
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