转基因克隆牛外源基因整合位点的研究  

Transgene Integration Site Analysis in Transgenic Bovine

作　　者：谭贝贝[1] 苏红[2] 胡嘉祺 吴茜红[1] 徐大庆[1] 巩校东[1] 李世杰[1] 

机构地区：[1]河北农业大学生命科学学院,河北保定071001 [2]中国地质大学长城学院,河北保定071000

出　　处：《中国农学通报》2012年第17期28-32,共5页Chinese Agricultural Science Bulletin

基　　金：转基因生物新品种培育重大专项"高通量基因克隆技术体系的研究"(2011ZX08009-002);国家重大科技专项(2008ZX08006-004);国家自然科学基金"体细胞克隆牛Dlk1-Gtl2印记区域表观遗传重编程的研究"(30972098)

摘　　要：整合位点的侧翼序列是外源基因表型研究和功能探讨的前提条件,对目的基因的正常转录和表达有着重要影响。本试验利用热不对称交措式PCR(Thermal Asymmetric Interlaced PCR,TAIL-PCR)的方法克隆了目的基因。结果表明:外源基因整合到牛的12号染色体基因组克隆(NW_003104315.1)中。分析了整合位点的纯合性发现转基因牛为整合位点杂合子。TAIL-PCR法能够有效、快速的鉴定外源基因在基因组中的整合位置,具有良好的应用前景。It was important to detect transgene integration site after establishment of transgenic animal line, and it was necessary to determine transgene expression and function. The integration site was successfully amplified using Thermal Asymmetric Interlaced PCR （TAIL-PCR）. The results showed that the target sequence was integrated into chromosome 12 （NW_003104315.1）. The homozygosis analysis of integration site showed the transgenic bovine was heterozygosity. The transplanted gene could be easily and quickly located in the genomic sequence and TAIL-PCR has advanced future application.

关 键 词：TAIL-PCR 转基因牛 整合位点 纯合性 

分 类 号：Q78[生物学—分子生物学]