慢病毒载体体外转染鸡胚原始生殖细胞  

Lentiviral Vector Transfection of Chicken Embryo Primordial Germ Cells In vitro

作　　者：丁红梅[1] 徐世永[1] 邵根宝[1] 孙岩[1] 王蒙[1] 刘红林[1] 

机构地区：[1]南京农业大学动物科技学院,南京210095

出　　处：《农业生物技术学报》2007年第4期612-616,共5页Journal of Agricultural Biotechnology

基　　金：国家948项目(No.202084)资助

摘　　要：提取第28期伊萨鸡(Gallus domesticus)生殖嵴原始生殖细胞(PGCs),将其与性腺基质细胞共培养,并对PGCs进行PAS(过碘酸希夫试剂)和AKP(碱性磷酸酶)染色鉴定。构建pL-CMV-eGFP慢病毒表达载体,与辅助质粒共转染293FT细胞生产病毒颗粒。将慢病毒浓缩后转染鸡胚PGCs,转染率高达24.19%。Primordial germ cells （PGCs） were isolated from the gonads of ISA brown chicken （Gallus domesticus） embryos at stage 28 and PGCs were co-cultured with gonadal stroma cells. Identification of PGCs was carried out by periodic acid-Schiff （PAS） and alkaline phosphatase （AKP） staining. A lentiviral vector pL-CMV-eGFP was constructed and harvested the virus by cotransfecting 293FT cells with the vector and packaging plasmids. Lentiviruses after concentration were used to transfect chicken PGCs, and the transfection efficiency of PGCs was up to 24.19%..

关 键 词：鸡胚 原始生殖细胞 慢病毒 转染效率 

分 类 号：S188[农业科学—农业基础科学]