摘要
根据GenBank中公布的牛、人等物种的Ghrelin核苷酸序列设计合成一对引物,以水牛皱胃组织总RNA为模板,采用RT-PCR法,扩增出Ghrelin成熟蛋白编码cDNA序列,将此扩增产物克隆入pMD18-T载体,进行PCR、双酶切鉴定及序列测定与分析。结果表明,扩增的水牛Ghre-lin基因编码序列长为351 bp,编码116个氨基酸。同源性分析表明,水牛Ghrelin基因与牛、人、猪、小鼠及绵羊基因相应序列的同源性分别为97%、79%、81%、75%和95%,氨基酸同源性与牛、羊、人、猪、小鼠分别为96%,94%,73%,76%和75%,说明Ghrelin是一组在进化上高度保守的蛋白质。水牛Ghrelin成熟蛋白cDNA的成功克隆,为进一步研究水牛Ghrelin基因结构、基因表达与调控奠定了基础。
A pair of primers was designed and synthesized according to the gene sequence of cow Ghrelin. RT-PCR method was used to amplify Ghrelin gene from the whole RNA of abomasum tissue in swamp buffalo. Sequence analysis revealed that the gene was 351 bp in length, encoding 116 amino acids and exhibited a high homology with those from cow （97%）, sheep （95%）, pig （81%）, mouse （75%） and human （73%）. The result indicated that Ghrelin gene is highly conservative in evolution. This study lays a foundation for further analysis of structure, expression and regulation of Ghrelin gene in swamp buffalo.
出处
《畜牧与兽医》
北大核心
2009年第1期25-28,共4页
Animal Husbandry & Veterinary Medicine
基金
国家"863"计划资助项目(2002AA06651)
