猪生长激素基因的克隆及在哺乳动物细胞中的表达  

Cloning and expression of porcine growth hormone gene in mammalian cells

作　　者：刘燕飞[1] 尉研[1] 付若彬[1] 丁秀琼[1] 谭俊杰[1] 孟亚鹏[1] 刘世贵[1] 龙章富[2] 

机构地区：[1]四川大学生命科学学院,四川成都610064 [2]四川大学生命科成都川大创新生物技术研究所,四川成都610064

出　　处：《中国兽医科技》2005年第2期134-138,共5页Chinese Journal of Veterinary Science and Technology

基　　金：教育部重点项目(国教00104);四川省"十五"重点项目(03NC002 001);四川省应用基础研究项目(02NY029 079)

摘　　要：应用RT-PCR技术克隆了猪生长激素(pGH)cDNA,该基因编码蛋白的信号肽序列与已有报道的pGH基因存在2个氨基酸残基的差异,而成熟肽却无差异。将 pGH cDNA定向插入真核表达载体VR1020,构建了重组真核表达质粒VpGH;利用脂质体法转染哺乳动物细胞COS7,对转染后的COS7细胞进行RT-PCR、ELISA和免疫荧光分析,分别在转录和翻译水平证实了目的基因在COS7细胞中得到正确转染表达。A cDNA for porcine growth hormone (pGH) was amplified by RT-PCR using RNA of porcine pituitary as the template. There were 2 amino acids difference in the signal peptide but no change in the mature peptide between new pGH and the other published reports of porcine GH was found. The recombinant expression vector was constructed by inserting the pGH cDNA into eukaryotic expression vector VR1020. Using lipofectin methods, the recombinant expression plasmid was transfected into COS7 cells. RT-PCR,ELISA and Immunofluorescence assay confirmed that pGH gene had been correctly transcripted and translated in COS7 cells.

关 键 词：猪生长激素基因 克隆 真核表达 

分 类 号：Q786[生物学—分子生物学]