人乳铁蛋白基因乳腺特异性表达载体的构建及转染研究  

Construction and identification of the mammary gland special expression vector for human lactoferritin

作　　者：张晶晶[1] 刘凤军[1] 彭淑英[1] 宋红卫[1] 张涌[1] 

机构地区：[1]西北农林科技大学生物工程研究所,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2007年第1期29-32,共4页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家"863"计划项目(2001AA213081)

摘　　要：构建了携带人乳铁蛋白(LTF)基因、绿色荧光蛋白(GFP)基因的乳腺特异性表达载体(pEBL),用脂质体法转染泌乳期莎能奶山羊乳腺上皮细胞,转染后于荧光显微镜下观察GFP表达情况。用G 418对转染细胞进行筛选,获得抗性细胞克隆,并对转染细胞进行PCR检测。结果显示,pEBL构建成功,转染6 h后可观察到细胞有GFP表达,PCR检测阳性克隆细胞转有LTF基因。表明pEBL载体已转染山羊乳腺上皮细胞,为进一步建立转基因动物乳腺生物反应器奠定了基础。The mammary gland special expression vector pEBL containing green fluorescence protein （GFP） and human lactoferritin （LTF） was constructed and transfected into goat mammary epithelial cells （GMEC） by liposome. Clones were screened by G418 and were identified by PCR. The GFP expressed cells were detected in 6 hours after transfection,PCR result showed that LTF gene was expressed in transfected cells. The vector pEBL was transfected successfully and efficiently. This lays a solid foundation for constructing transgenic animals and its mammary gland hioreactor.

关 键 词：山羊乳腺上皮细胞 人乳铁蛋白 绿色荧光蛋白 特异性载体 

分 类 号：Q782[生物学—分子生物学]