广西巴马小型猪生长素(Ghrelin)cDNA的克隆及序列分析

目的扩增广西巴马小型猪生长素(Ghrelin)cDNA序列,分析其序列结构特点,为进一步从分子水平上对广西巴马小型猪的生长发育提供理论依据,也为日后构建原核表达载体、研究Ghrelin基因的生物功能奠定基础。方法以广西巴马小型猪胃底组织总RNA为模板,用特异性引物扩增Ghrelin的cDNA,纯化连接pMD18-T载体后转化大肠杆菌DH5α,筛选阳性克隆体,鉴定后测序。结果经测序证实克隆得到的广西巴马小型猪Ghrelin cDNA序列全长为357 bp,与相关文献报道一致。同源性比较发现,广西巴马小型猪Ghrelin cDNA基因序列与GenBank报道的猪相应序列同源性为99.7%,只有一处发生碱基突变。结论本实验成功克隆了广西巴马小型猪Ghrelin cDNA。

Cloning and Sequence Analysis of Ghrelin cDNA from Guangxi Bama Minipig

Objective To clone and analyze the sequence of Ghrelin cDNA of Guangxi Bama minipig. Methods The sequence of Ghrelin eDNA was amplified by a specific pair of primers using Guangxi Bama minipig belly total RNA as template. The Ghrelin eDNA was cloned by RT-PCR and then ligated into pMD18-T vector after purification. The recombined pMD18-T vector was transformed into the coliform DH5a for sequencing. Results The cloned Ghrelin eDNA gene from Guangxi Bama minipig contained 357 bp. The homology analysis showed that compared with the. sequence of other pig breeds reported in GenBank, the homology rate of the Ghrelin eDNA sequence were 99.7 %. Conclusion Sequencing confirmed that the sequence of Ghrelin eDNA of Guangxi Bama minipig was successfully cloned in the study.