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马传染性贫血病毒慢病毒逆转录酶的表达及突变分析

Expression and mutational analysis of the reverse transcriptase of the lentivirus equine infectious anemia virus.

作者信息

Shaharabany M, Rice N R, Hizi A

机构信息

Department of Cell Biology and Histology, Sackler School of Medicine, Tel Aviv University, Israel.

出版信息

Biochem Biophys Res Commun. 1993 Oct 29;196(2):914-20. doi: 10.1006/bbrc.1993.2336.

Abstract

The reverse transcriptase of equine infectious anemia virus (EIAV) shows sequence similarity with the reverse transcriptases of other lentiviruses, particularly with those of human immunodeficiency viruses types 1 and 2 (HIV-1 and HIV-2). We have constructed a plasmid that when introduced into E. coli induces the synthesis of substantial quantities of the nearly authentic EIAV reverse transcriptase. The viral and bacterially expressed reverse transcriptases are similar in their molecular weights. The bacterial expression clone was used to generate deletion mutants of the protein. Mutations in both amino and carboxyl terminal regions of the polypeptide strongly affect the DNA polymerase activity of the enzyme. Thus, EIAV reverse transcriptase resembles the reverse transcriptases of HIV-1 and HIV-2 and can serve as a suitable enzyme for studying the structure-function relationship in lentiviral reverse transcriptase.

摘要

马传染性贫血病毒(EIAV)的逆转录酶与其他慢病毒的逆转录酶在序列上具有相似性,尤其是与1型和2型人类免疫缺陷病毒(HIV-1和HIV-2)的逆转录酶。我们构建了一种质粒,将其导入大肠杆菌后可诱导合成大量近乎天然的EIAV逆转录酶。病毒表达的逆转录酶和细菌表达的逆转录酶分子量相似。利用细菌表达克隆产生该蛋白的缺失突变体。多肽氨基末端和羧基末端区域的突变均强烈影响该酶的DNA聚合酶活性。因此,EIAV逆转录酶与HIV-1和HIV-2的逆转录酶相似,可作为研究慢病毒逆转录酶结构-功能关系的合适酶。

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