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豚鼠巨细胞病毒糖蛋白B基因的克隆与鉴定

Cloning and characterization of the guinea pig cytomegalovirus glycoprotein B gene.

作者信息

Schleiss M R

机构信息

Division of Infectious Diseases, Children's Hospital Research Foundation, Cincinnati, Ohio 45229.

出版信息

Virology. 1994 Jul;202(1):173-85. doi: 10.1006/viro.1994.1333.

DOI:10.1006/viro.1994.1333
PMID:8009831
Abstract

Unlike other small animals, cytomegalovirus (CMV) infection of the guinea pig results in transplacental passage and intrauterine infection of the fetus. These features make the guinea pig model ideal for studying vaccine strategies designed to prevent congenital infection. Unfortunately, little is known about immunogenic guinea pig CMV gene products. In other animal cytomegaloviruses, a major target of the host immune response is the glycoprotein B (gB, gp UL 55) gene product. Using DNA probes containing human CMV gB sequences, the gB gene homolog of the guinea pig cytomegalovirus was identified, cloned, and sequenced. The gpCMV gB gene maps to a region spanning portions of the HindIII K, Q, and P fragments of the gpCMV genome. DNA sequence analysis identified an open reading frame of 2706 nucleotides capable of encoding a protein of 901 amino acids. Extensive similarity to the human and murine gB proteins was noted with 42% identity at the amino acid level. The predominant gpCMV gB mRNA is a 6.8-kb transcript with the expression kinetics of an early gene. RNase protection and primer extension analyses indicated that gB mRNAs were transcribed from two different initiation sites corresponding to distinct TATA elements. Polyclonal antisera prepared against a synthetic peptide derived from amino acid sequences within the ORF identified a 58-kDa virion-associated protein representing the cleaved COOH-terminus (gp 58) of the gpCMV gB molecule. The molecular characterization of gpCMV gB should facilitate studies of vaccine strategies in the guinea pig model of congenital CMV infection.

摘要

与其他小型动物不同,豚鼠感染巨细胞病毒(CMV)会导致病毒经胎盘传播并感染胎儿子宫内的胎儿。这些特征使豚鼠模型成为研究旨在预防先天性感染的疫苗策略的理想选择。不幸的是,关于具有免疫原性的豚鼠CMV基因产物知之甚少。在其他动物巨细胞病毒中,宿主免疫反应的主要靶标是糖蛋白B(gB,gp UL 55)基因产物。使用含有人类CMV gB序列的DNA探针,鉴定、克隆并测序了豚鼠巨细胞病毒的gB基因同源物。gpCMV gB基因定位于跨越gpCMV基因组HindIII K、Q和P片段部分的区域。DNA序列分析确定了一个2706个核苷酸的开放阅读框,能够编码一个901个氨基酸的蛋白质。在氨基酸水平上,与人类和鼠类gB蛋白有广泛的相似性,同一性为42%。主要的gpCMV gB mRNA是一个6.8 kb的转录本,具有早期基因的表达动力学。核糖核酸酶保护和引物延伸分析表明,gB mRNA从两个不同的起始位点转录,对应于不同的TATA元件。针对从开放阅读框内氨基酸序列衍生的合成肽制备的多克隆抗血清,鉴定出一种58 kDa的病毒体相关蛋白,代表gpCMV gB分子的切割COOH末端(gp 58)。gpCMV gB的分子特征应有助于在先天性CMV感染的豚鼠模型中研究疫苗策略。

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