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猴水痘病毒糖蛋白B基因的DNA序列及转录分析

DNA sequence and transcriptional analysis of the simian varicella virus glycoprotein B gene.

作者信息

Pumphrey C Y, Gray W L

机构信息

Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock 72205.

出版信息

J Gen Virol. 1994 Nov;75 ( Pt 11):3219-27. doi: 10.1099/0022-1317-75-11-3219.

Abstract

The varicella-zoster virus (VZV) glycoprotein B (gB) is a major viral antigen which elicits immunity and neutralizing antibodies. In this study, the genomic map position and DNA sequence of a simian varicella virus (SVV) homologue of the VZV gB gene was identified and the transcript analysed. A 32P-labelled VZV gB DNA probe hybridized to a subclone of the SVV BamHI B restriction endonuclease fragment indicating the fine map position of SVV DNA sequences homologous to the VZV gB gene. The SVV gB DNA sequence was determined and analysis revealed a 2751 base pair open reading frame (ORF) with 71.1% identity to the VZV gB gene and 53.8% identity to the herpes simplex type 1 gB gene. The SVV gB ORF encodes a 916 amino acid polypeptide with a predicted molecular mass of 104K. The deduced SVV and VZV gB polypeptides share 78.9% amino acid identity and predicted N-linked glycosylation sites, cleavage sites and transmembrane regions. 32P-labelled SVV gB DNA and RNA probes hybridized to a 3.5 kilobase SVV polyadenylated transcript. Primer extension experiments identified transcript start sites for the SVV and VZV gB genes and permitted a comparison of the sequences upstream of the SVV and VZV gB ORFs. The SVV and VZV gB promoter elements are similar in content and align closely. The VZV gB transcript start site suggests a gB polypeptide initiation site which is inconsistent with the previously reported ATG start codon.

摘要

水痘带状疱疹病毒(VZV)糖蛋白B(gB)是一种主要的病毒抗原,可引发免疫反应和中和抗体。在本研究中,确定了VZV gB基因的猿猴水痘病毒(SVV)同源物的基因组图谱位置和DNA序列,并对转录本进行了分析。一个用32P标记的VZV gB DNA探针与SVV BamHI B限制性内切酶片段的一个亚克隆杂交,表明与VZV gB基因同源的SVV DNA序列的精细图谱位置。测定了SVV gB DNA序列,分析显示有一个2751个碱基对的开放阅读框(ORF),与VZV gB基因的同一性为71.1%,与单纯疱疹病毒1型gB基因的同一性为53.8%。SVV gB ORF编码一个916个氨基酸的多肽,预测分子量为104K。推导的SVV和VZV gB多肽具有78.9%的氨基酸同一性以及预测的N-连接糖基化位点、切割位点和跨膜区域。用32P标记的SVV gB DNA和RNA探针与一个3.5千碱基的SVV多聚腺苷酸化转录本杂交。引物延伸实验确定了SVV和VZV gB基因的转录起始位点,并允许对SVV和VZV gB ORF上游的序列进行比较。SVV和VZV gB启动子元件在内容上相似且排列紧密。VZV gB转录起始位点提示了一个gB多肽起始位点,这与先前报道的ATG起始密码子不一致。

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