在大肠杆菌中筛选靶向人类免疫缺陷病毒（HIV）逆转录酶的核苷类似物：HIV逆转录酶与单纯疱疹病毒胸苷激酶的共表达
A screen in Escherichia coli for nucleoside analogs that target human immunodeficiency virus (HIV) reverse transcriptase: coexpression of HIV reverse transcriptase and herpes simplex virus thymidine kinase.
作者信息
Kim B, Loeb L A
机构信息
Joseph Gottstein Memorial Cancer Research Laboratory, Department of Pathology and Biochemistry, University of Washington, Seattle 98195, USA.
出版信息
J Virol. 1995 Oct;69(10):6563-6. doi: 10.1128/JVI.69.10.6563-6566.1995.
Abstract
Human immunodeficiency virus (HIV) reverse transcriptase substitutes for temperature-sensitive DNA polymerase I (Pol Its) in Escherichia coli, providing a screen for anti-HIV reverse transcriptase nucleoside analogs in bacteria. Since phosphorylation of nucleosides in E. coli is limited to thymidine and its derivatives, we coexpressed herpes simplex virus thymidine kinase, an enzyme that phosphorylates a wide variety of nucleoside analogs, together with HIV reverse transcriptase. Coexpression of herpes simplex virus thymidine kinase and HIV reverse transcriptase rendered Pol Its cells sensitive to dideoxycytidine. Studies with different nucleoside analogs indicate that this bacterial screening system is able to select and identify nucleoside analogs that specifically target HIV reverse transcriptase.
摘要
人类免疫缺陷病毒（HIV）逆转录酶可替代大肠杆菌中温度敏感型DNA聚合酶I（Pol Its），为在细菌中筛选抗HIV逆转录酶核苷类似物提供了一种方法。由于大肠杆菌中核苷的磷酸化仅限于胸苷及其衍生物，我们将单纯疱疹病毒胸苷激酶（一种能使多种核苷类似物磷酸化的酶）与HIV逆转录酶共表达。单纯疱疹病毒胸苷激酶和HIV逆转录酶的共表达使Pol Its细胞对双脱氧胞苷敏感。对不同核苷类似物的研究表明，这种细菌筛选系统能够选择和鉴定特异性靶向HIV逆转录酶的核苷类似物。
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