限制性显示技术制备HIV-1B亚型基因片段的研究  

Preparation of Gene Fragments from HIV-1 U26942 by Restriction Display

作　　者：李凌[1] 马文丽[1] 甄莉[2] 吴清华[1] 刘翠华[1] 郑文岭[3] 

机构地区：[1]第一军医大学分子生物学研究所 [2]南方医院,广州510515 [3]广州总医院分子肿瘤学研究所,广州510010

出　　处：《病毒学报》2002年第2期177-180,共4页Chinese Journal of Virology

基　　金：广东省自然科学基金 (9840 92 ) ;广州市重点科技攻关项目 (990 44 80 2 2 )

摘　　要：In order to isolate expeditiously the HIV1U26942 DNA fragments for preparation of DNA microarrays, the multiple gene fragments with sizes suitable for DNA microarrays, produced by digesting the dissociated HIV gene with Sau 3AⅠ, were ligated with universal adapters. PCR primers were designed to match the universal adapters (including the restriction site sequence) but with one "nesting" base overhanging at the 3’ end. The PCR reactions that were performed with various single primers or primer combinations were divided into ten subgroups. PCR products were purified and then cloned into the T vectors. The positive clones were propagated and the plasmids were extracted. The target HIV gene fragments were isolated and sequenced, which were correlated precisely with the prediction of restriction analysis. Eighteen gene fragments ranging from 0.1kb to 1kb were prepared for DNA microarray. Restriction display is an effective and rapid method for the isolation of geneIn order to isolate expeditiously the HIV1U26942 DNA fragments for preparation of DNA microarrays, the multiple gene fragments with sizes suitable for DNA microarrays, produced by digesting the dissociated HIV gene with Sau 3AⅠ, were ligated with universal adapters. PCR primers were designed to match the universal adapters (including the restriction site sequence) but with one 'nesting' base overhanging at the 3' end. The PCR reactions that were performed with various single primers or primer combinations were divided into ten subgroups. PCR products were purified and then cloned into the T vectors. The positive clones were propagated and the plasmids were extracted. The target HIV gene fragments were isolated and sequenced, which were correlated precisely with the prediction of restriction analysis. Eighteen gene fragments ranging from 0.1kb to 1kb were prepared for DNA microarray. Restriction display is an effective and rapid method for the isolation of gene fragments

关 键 词：限制性显示技术 制备 HIV-1B亚型 基因片段 人类免疫缺陷病毒1型 DNA芯片 

分 类 号：R373.9[医药卫生—病原生物学] R512.91[医药卫生—基础医学]