绵羊乳腺特异表达人肝细胞再生增强因子载体的构建及表达

用PCR技术分别从人和绵羊基因组DNA中扩增人肝细胞再生增强因子(Human augmenter of liver regeneration，hALR)基因和绵羊β-乳球蛋白(sheep beta-lactoglobulin，BLG)基因启动区序列，从pEGFP-C1质粒中扩增增强绿色荧光蛋白(Enhanced Green fluorescence protein，EGFP)基因及其表达调控元件．由质粒p7zf( )构建成ALR基因乳腺特异表达、EGFP基因非组织特异性表达载体．同时体外培养绵羊胎儿成纤维细胞(sheep fetal fibroblast cells，SFFCs)，脂质体介导载体DNA转染SFFCs，激光共聚焦显微镜观察和PCR检测转基因细胞，结果表明，增强绿色荧光蛋白基因在SFFCs中表达，转基因细胞中可扩增出BLG、ALR、EGFP基因条带．

Construction and Expression of a Vector to Express ALR in Sheep Mammary Glands

Human ALR gene and sheep BLG genes promoter sequences were amplified from human and sheep genomic DNA using PCR.The breast specific expression of ALR constructed by plasmid p7zf(+) was controlled by BLG promoter,and the non-specific expression of EGFP gene was controlled by CMV dual promoter system as genetic marker in the same vector.Sheep fetal fibroblast cell were transfected with p7zf-BAE by using LipofectAMINETM substance.Transgenic cells were identified by fluorescent microscope and PCR technique.The results showed that the EGFP gene expressed independently in the SFFCs,and the BLG and ALR genes existed in the transgenic cells as well.