转座介导的HIV-1整合位点分析

Analysis of transposon-mediated HIV-1 integration sites

目的 建立转座酶反应介导的HIV-1整合位点鉴定方法,以鉴定HIV-1潜伏或复制性感染有关的整合特征.方法 采用转座酶反应介导整合序列的捕获与扩增方法,对照人基因组进行序列分析.结果 50株细胞克隆中,有45株可检测到HIV-1整合片段,共获得9个整合位点.HIV-1潜伏或复制型细胞内整合位点的分布无显著差异.HIV-1的整合位点分布于所有基因目录内,包括转录活跃基因、非编码区或重复元件,并与相邻的表观分布相关.结论 转座酶反应介导的整合位点鉴定方法简便快捷,能以最大随机方式还原整合位点.HIV-1感染的整合与基因组特征存在联系.

Objective To establish the identification method for transposon-mediated HIV-1 integration sites,so as to identify the integration characteristics of replicative or latent HIV-1 infection.Methods The integrated sequences were captured and amplified by the transposase reaction,and then further analyzed by mapping to human genome.Results Of 50 clones sequenced,45 clones had HIV-1 integration events with 9 integration sites determined by sequence analysis.Integration sites from HIV-1 were compared in latent or replicative T cell lines,which showed no significant difference.Integration sites of HIV-1 were distributed in almost all gene catalogues,including active transcribed genes,non-coding region and repetitive elements.Epigenomic features were associated with integration sites.Conclusions The integration site identification method mediated by transposase reaction is simple and rapid,which can recover the integration sites efficiently.The integration of HIV-1 infection is related to the genomic characteristics.