体内精原干细胞介导法制备抗病转基因鸡  

Generation of Antiviral Transgenic Chicken Using Spermatogonial Stem Cell in Vivo Transfection

作　　者：孙敏[1] 施青青[1] 阴彦辉[1] 傅德智[1] 秦玉蓉[1] 许峰[1] 李碧春[1] 

机构地区：[1]扬州大学动物科学与技术学院,江苏省动物繁育与分子设计重点实验室,江苏扬州225009

出　　处：《中国畜牧杂志》2011年第19期1-6,共6页Chinese Journal of Animal Science

基　　金：国家自然科学基金(30871791);“六大人才高峰”项目;江苏省高校自然科学重大基础研究项目(08KJA230001)

摘　　要：实验旨在利用精原干细胞介导的方法体内转染融合基因EGFP-MMx,制备抗病毒性疾病转基因鸡。采用脂质体包埋法,睾丸内注射EGFP-MMx融合基因,术后采集30、40、50、60、70、80 d睾丸组织进行冰冻切片、基因组PC R、点杂交实验,转染后60 d公鸡精液人工授精于正常母鸡,对后代血液采用PC R检测技术、R T-PC R和W estern blot方法检测外源基因EGFP-MMx的整合表达情况。结果表明:睾丸基因组点杂交阳性率为72.2%。通过PC R、R T-PCR检测方法,在实验公鸡精液和F1代血液中检测到目的基因,精液阳性率为25%,F1代阳性率为10.53%,Western blot检测F1代阳性率为7.89%。结果证明外源基因通过睾丸注射法可整合到基因组上,为培育抗病转基因鸡提供参考依据。In order to cultivate anti-viral transgenic chickens through transfected spermatogonial stem cell with fusion gene EGFP-MMx. After injecting fusion gene EGFP-MMx into testes, we did some testing such as frozen section, PCR and dot blot of testes＇ tissues of 30 d, 40 d, 50 d, 60 d, 70 d and 80 d, a few of normal hen were inseminated artificially with cock semen of 60 d. Finally we detected the integration of exogenous gene EGFP-MMx of the offspring by PCR, RT-PCR and Western Blot. Dot blot positive rate of testes genome was 72.2%. PCR and RT-PCR positive rate was 25% and 10.53%（4/38 ）in sperm and F1 blood genome respectively, while Western Blot positive rate of F1 blood was 7.89%（3/38 ）. It was proved the possibility of cultivating anti-viral transgenic chickens by the method of testes injection method which can integrate foreign genes into the genome.

关 键 词：精原干细胞 EGFP—MMx融合蛋白 转基因鸡 

分 类 号：S831.2[农业科学—畜牧学]