【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。更多还原

【Abstract】 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.更多还原