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马疱疹病毒1型UL1基因产物的鉴定与表达

Identification and expression of the UL1 gene product of equine herpesvirus 1.

作者信息

Harty R N, O'Callaghan D J

机构信息

Department of Microbiology and Immunology, Louisiana State University Medical Center, Shreveport 71130-3932.

出版信息

Virus Res. 1992 Sep 1;25(1-2):105-16. doi: 10.1016/0168-1702(92)90103-g.

Abstract

Sequences encoding the UL1 gene of equine herpesvirus type 1 (EHV-1) are conserved in the genome of EHV-1 defective interfering particles (DIPs) that mediate oncogenic transformation and persistent infection. The UL1 protein was identified by in vitro transcription/translation and hybrid-arrest translation analyses which employed a UL1/pGEM-3Z construct designated pGEML1. SDS-PAGE analyses of in vitro translation products synthesized from UL1-specific RNA revealed that the UL1 ORF encodes a 30 kDa protein which corresponds in size to the 258 amino acid protein predicted by DNA sequence analyses. This result was confirmed by arresting translation of the in vitro transcribed UL1 RNA with an oligodeoxynucleotide complementary to UL1 coding sequences. The UL1 protein is a homolog of the predicted protein encoded by the ORF2 gene of varicella-zoster virus, but UL1 has no homolog in herpes simplex virus type 1. The UL1 protein contains a region conforming to a 'PEST' (Proline, Glutamic acid, Serine, and Threonine) sequence, which is commonly found in proteins with half-lives of less than two hours.

摘要

编码1型马疱疹病毒(EHV-1)UL1基因的序列在介导致癌转化和持续感染的EHV-1缺陷干扰颗粒(DIPs)基因组中是保守的。通过体外转录/翻译和杂交阻断翻译分析鉴定了UL1蛋白,这些分析采用了一个名为pGEML1的UL1/pGEM-3Z构建体。对由UL1特异性RNA合成的体外翻译产物进行的SDS-PAGE分析表明,UL1开放阅读框编码一种30 kDa的蛋白质,其大小与DNA序列分析预测的258个氨基酸的蛋白质一致。用与UL1编码序列互补的寡脱氧核苷酸阻断体外转录的UL1 RNA的翻译,证实了这一结果。UL1蛋白是水痘带状疱疹病毒ORF2基因编码的预测蛋白的同源物,但UL1在1型单纯疱疹病毒中没有同源物。UL1蛋白包含一个符合“PEST”(脯氨酸、谷氨酸、丝氨酸和苏氨酸)序列的区域,该序列常见于半衰期小于两小时的蛋白质中。

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