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大鼠巨细胞病毒R32基因编码一种病毒体相关蛋白,该蛋白在受感染的大鼠中引发强烈的体液免疫反应。

The rat cytomegalovirus R32 gene encodes a virion-associated protein that elicits a strong humoral immune response in infected rats.

作者信息

Beuken Erik, Grauls Gert, Bruggeman Cathrien A, Vink Cornelis

机构信息

Department of Medical Microbiology, Cardiovascular Research Institute Maastricht, Maastricht University, PO Box 5800, 6202 AZ Maastricht, The Netherlands1.

出版信息

J Gen Virol. 1999 Oct;80 ( Pt 10):2719-2728. doi: 10.1099/0022-1317-80-10-2719.

Abstract

A gene of rat cytomegalovirus (RCMV), designated R32, has been identified that encodes a homologue of the human cytomegalovirus (HCMV) pp150 (ppUL32) major tegument phosphoprotein. The R32 ORF has the capacity to encode a 667 amino acid polypeptide (pR32) with a calculated molecular mass of 73 kDa. The predicted amino acid sequence of pR32 shows similarity to that of polypeptides predicted to be encoded by the HCMV UL32, murine cytomegalovirus M32 and human herpesvirus types 6 and 7 U11 genes. The R32 gene is transcribed as a 2.5 kb mRNA during the late phase of RCMV infection in rat embryo fibroblasts in vitro. To study expression of the pR32 protein in vitro and in vivo, a rabbit polyclonal antiserum was raised against a recombinant protein that comprised amino acids 252-522 of pR32. By using this antiserum, pR32 could be detected predominantly in the cytoplasm of RCMV-infected fibroblasts at 24 and 48 h post-infection in vitro. The pR32 protein was also detected within virions isolated from the culture medium of RCMV-infected cells. Expression of pR32 in vivo was observed within the cytoplasm of salivary gland epithelial cells of RCMV-infected rats. In addition, recombinant pR32 was found to react with sera from rats that were previously infected with RCMV, whereas reactivity was not seen with sera from mock-infected rats. Together, these findings indicate that RCMV pR32 represents the homologue of HCMV ppUL32, both in primary structure and in function.

摘要

已鉴定出大鼠巨细胞病毒(RCMV)的一个名为R32的基因,它编码人巨细胞病毒(HCMV)pp150(ppUL32)主要衣壳磷蛋白的同源物。R32开放阅读框能够编码一个667个氨基酸的多肽(pR32),计算分子量为73 kDa。预测的pR32氨基酸序列与预测由HCMV UL32、小鼠巨细胞病毒M32以及人疱疹病毒6型和7型U11基因编码的多肽序列相似。在体外大鼠胚胎成纤维细胞中RCMV感染的晚期,R32基因转录为2.5 kb的mRNA。为了研究pR32蛋白在体外和体内的表达,制备了针对包含pR32第252 - 522位氨基酸的重组蛋白的兔多克隆抗血清。使用该抗血清,在体外感染后24小时和48小时,pR32主要在RCMV感染的成纤维细胞的细胞质中被检测到。从RCMV感染细胞的培养基中分离的病毒粒子内也检测到了pR32蛋白。在RCMV感染大鼠的唾液腺上皮细胞的细胞质中观察到了pR32在体内的表达。此外,发现重组pR32与先前感染RCMV的大鼠的血清发生反应,而与模拟感染大鼠的血清未观察到反应性。总之,这些发现表明RCMV pR32在一级结构和功能上均代表HCMV ppUL32的同源物。

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