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开发一种基于酵母的重组克隆系统,用于分析来自多种1型人类免疫缺陷病毒分离株的基因产物。

Development of a yeast-based recombination cloning/system for the analysis of gene products from diverse human immunodeficiency virus type 1 isolates.

作者信息

Marozsan Andre J, Arts Eric J

机构信息

Division of Infectious Diseases, Department of Medicine, Case Western Reserve University, BRB 1029, 10900 Euclid Avenue, Cleveland, OH 44106, USA.

出版信息

J Virol Methods. 2003 Aug;111(2):111-20. doi: 10.1016/s0166-0934(03)00166-6.

Abstract

A recent shift from studies on a few subtype B laboratory human immunodeficiency virus type 1 (HIV-1) clones to analyses of extremely diverse primary HIV-1 isolates from different subtype requires the development of a rapid and generic cloning technique. This report describes the use of gap repair/recombination in yeast to shuttle env, gag, and pol genes from diverse HIV-1 subtypes into a DNA vector that can be amplified in bacteria and can express the gene of interest in mammalian cells. These diverse HIV-1 genes have also been introduced into an infectious clone to produce chimeric viruses that are useful for studies on drug susceptibility, receptor binding and fitness.

摘要

最近,研究已从对少数B亚型实验室1型人类免疫缺陷病毒(HIV-1)克隆的研究转向对来自不同亚型的极其多样的原发性HIV-1分离株的分析,这就需要开发一种快速且通用的克隆技术。本报告描述了利用酵母中的缺口修复/重组将来自不同HIV-1亚型的env、gag和pol基因穿梭到一个DNA载体中,该载体可在细菌中扩增并能在哺乳动物细胞中表达感兴趣的基因。这些多样的HIV-1基因也已被引入到一个感染性克隆中,以产生嵌合病毒,这些病毒可用于药物敏感性、受体结合和适应性研究。

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