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一种编码膜蛋白的人巨细胞病毒基因的鉴定与特性分析,该膜蛋白在人疱疹病毒中保守。

Identification and characterization of a human cytomegalovirus gene coding for a membrane protein that is conserved among human herpesviruses.

作者信息

Lehner R, Meyer H, Mach M

机构信息

Institut für Klinische und Molekulare Virologie, Universität Erlangen-Nürnberg, Federal Republic of Germany.

出版信息

J Virol. 1989 Sep;63(9):3792-800. doi: 10.1128/JVI.63.9.3792-3800.1989.

Abstract

A rabbit antiserum was raised against envelope material from purified human cytomegalovirus strain AD169. The serum recognized polypeptides 200, 170, 160, 75, 58, and 45 kilodaltons in size. It was used to screen a cDNA library constructed from poly(A)+ RNA from human cytomegalovirus-infected cells in the expression vector lambda gt11. A recombinant bacteriophage expressing cytomegalovirus-specific sequences was identified, and the corresponding gene was mapped to the HindIII R fragment. The gene is transcribed into a late 1.5-kilobase RNA. The nucleotide sequence of the coding region was determined. Computer analysis of the gene product revealed a polypeptide containing multiple potential membrane-spanning domains, representing a type of protein not identified in the envelope of herpesviruses before. The protein shows homology on the amino acid level to hypothetical proteins from reading frames BBRF3 of Epstein-Barr virus, UL10 of herpes simplex virus type 1, and ORF50 of varicella-zoster virus. By using an antiserum raised against procaryote-expressed parts of the cytomegalovirus membrane protein, a 45-kilodalton structural component of the virus was identified as the gene product.

摘要

制备了针对纯化的人巨细胞病毒AD169株包膜成分的兔抗血清。该血清识别大小为200、170、160、75、58和45千道尔顿的多肽。用它筛选了一个由人巨细胞病毒感染细胞的聚腺苷酸加尾RNA构建的、插入表达载体λgt11的cDNA文库。鉴定出一个表达巨细胞病毒特异性序列的重组噬菌体,并将相应基因定位到HindIII R片段。该基因转录成一种1.5千碱基的晚期RNA。测定了编码区的核苷酸序列。对该基因产物的计算机分析显示,其多肽含有多个潜在的跨膜结构域,代表一种以前在疱疹病毒包膜中未鉴定出的蛋白质类型。该蛋白质在氨基酸水平上与爱泼斯坦-巴尔病毒的阅读框BBRF3、单纯疱疹病毒1型的UL10以及水痘-带状疱疹病毒的ORF50中的假定蛋白质具有同源性。通过使用针对原核表达的巨细胞病毒膜蛋白部分产生的抗血清,鉴定出病毒的一种45千道尔顿的结构成分是该基因产物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f95e/250972/43329279e3c1/jvirol00076-0250-a.jpg

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