A lentiviral cDNA library employing lambda recombination used to clone an inhibitor of human immunodeficiency virus type 1-induced cell death
A lentiviral cDNA library employing lambda recombination used to clone an inhibitor of human immunodeficiency virus type 1-induced cell death
复制标题
DOI：
10.1128/jvi.78.20.11352-11359.2004
复制
发表时间：
2004-10-01
影响因子：
5.4
通讯作者：
Koyanagi, Y
中科院分区：
文献类型：
--
作者：
Kawano, Y;Yoshida, T;Koyanagi, Y
cdna表达克隆技术是鉴定基因新功能特性的合适工具。在这里，我们基于噬菌体lambda的位点特异性重组系统为人类T细胞生成了一个慢病毒cDNA文库表达系统，用于以最小的复杂性损失转移cDNA文库。用野生型人类免疫缺陷病毒1型（HIV-1）攻击文库转导的CD4(+) T细胞，选择对HIV-1诱导的细胞病变效应（CPE）产生抗性的细胞。从这些细胞中分离出CD14，并证明其抑制HIV-1和HIV-1诱导的CPE的进入。这种克隆系统可以快速识别人类T细胞和其他哺乳动物细胞中编码新特性的基因。
Expression cloning technology of cDNAs is a suitable tool for identifying novel functional properties of genes. Here, we generated a lentiviral cDNA library-expressing system for human T cells based on a site-specific recombination system of phage lambda for transferring cDNA libraries with a minimum loss of its complexity. The library-transduced CD4(+) T cells were challenged with wild-type human immunodeficiency virus type 1 (HIV-1), and the cells that acquired resistance to HIV-1-induced cytopathic effect (CPE) were selected. From these cells, CD14 was isolated and proved to inhibit the entry of HIV-1 and the HIV-1-induced CPE. This cloning system allows rapid identification of genes encoding novel properties in human T cells and probably other mammalian cells.