中文摘要：

本研究旨在建立一个用于干细胞示踪的报告体系，将EGFPcDNA及两侧带有LoxP位点的neo抗性基因插入五指山小型猪内源性Oct-4基因的终止密码子处，以Oct-4基因完整的51调控区启动EGFP的表达，为猪干细胞研究提供有价值的工具。试验定制了针对猪Oct-4终止密码子的TALENs，与打靶载体共转染猪耳成纤维细胞，药物筛选得到抗性克隆点514个，经过PCR鉴定，共获得杂合阳性克隆点36个，打靶效率分别为5．6％和13．0％。本研究成功获得了Oct-4-EGFP转基因细咆系，并证明了TALENs技术可以明显提高同源重组效率。

英文摘要：

This study aims to establish a reporting system for stem cells tracer, EGFP cDNA together with LoxP-flanked PGK-neo cassettes were inseet into the Oct-4 gene of Wuzhishan Miniature pig at the stop codon via TALENs mediated homolo- gous recombination, expression of EGFP gene was under control of Oct-4 5＇-regulatory region. TALENs directed against se- quences flanking the stop eodon of Oct 4 were cotrasfected with targeting vector, 514 drug-resistant cell clones were screened, and 36 correctly targeted clones were confirmed by PCR from them, the targeting efficiency were 5.6% and 13.0%. Oct-4- EGFP transgenic cell line was establinshed in this study, and it was also proved that TALENs could significantly improve the efficiency of homologous recombination.