转基因克隆牛外源基因整合位点的研究

Transgene Integration Site Analysis in Transgenic Bovine

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.