贵州香猪生长激素cDNA的克隆及原核表达  

Consturction and prokaryotic expression of the cDNA of growth hormone from Xiang pig

作　　者：李景[1] 冉雪琴[2] 王嘉福[1] 

机构地区：[1]贵州大学动物科学学院,550025 [2]贵州大学农业生物工程省重点实验室

出　　处：《贵州畜牧兽医》2007年第6期1-3,共3页Guizhou Journal of Animal Husbandry & Veterinary Medicine

基　　金：贵州省优秀青年科技人才培养计划(黔科合人字(2005)0506号);贵州省"十五"农业科技计划攻关(黔科合2004NGY010);贵州省"十一.五"农业科技重大专项(黔科合NZ字[2005]3002)

摘　　要：应用RT-PCR技术从香猪垂体mRNA中扩增出完整的生长激素(growth hormone,GH)cDNA片段,将其克隆入载体质粒pTYB11中,构建了重组质粒pTYB11-xGH,经过IPTG诱导,在大肠杆菌ER2566中获得表达,并确定了最佳表达条件。通过亲合柱层析,得到了完整的香猪生长激素,为香猪GH的结构、功能及其分子水平的作用机制奠定了基础。Growth hormone （GH） is a major regulator of growth, somatic development and body composition. GH eDNA was amplified from the pituitary total RNA of Xiang pig by RT - PCR and was cloned into a recombinant plasmid designed to be pMD - 18 - T - GH. The GH eDNA of Xiang pig contained an open reading frame encoding 216 amino acids of preprotein. The deduced mature peptide at position 108 was Ile. The coding DNA for mature GH of Xiang pig was amplified using pMD - 18 - T - GH as template to construct the recombinant plasmid, pTYB11 - xGH. The plasmid pTYB11 - xGH was induced by IPTG to produce fusion protein with mature GH of Xiang pig fused at the C - terminus of intein. After induced by DTT, the mature GH was further purified using chitin column. It suggested that the special GH with substitution of Ile at position 108 from Xiang pig would be a useful tool for deciphering the role of GH mechanism and function.

关 键 词：生长激素 定点突变 蛋白表达 

分 类 号：Q785[生物学—分子生物学]