广西巴马小型猪促生长激素释放激素成熟肽的克隆及序列分析

目的扩增广西巴马小型猪促生长激素释放激素(GHRH),分析其序列结构特点。方法以广西巴马小型猪基因组DNA为模板,用特异性引物扩增GHRH外显子3,再用引物延伸悬挂PCR法扩增GHRH成熟肽,纯化后,连接、转化到大肠杆菌,筛选阳性克隆,测序。结果经测序证实广西巴马小型猪GHRH外显子3及其成熟肽的基因序列与相关文献报道一致。克隆得到的广西巴马小型猪GHRH序列全长为132 bp,其中外显子3基因序列长105bp。同源性比较发现,广西巴马小型猪GHRH外显子3基因序列与GenBank(NCBI)中报到的猪GHRH外显子3相比,同源性为97.14%,有3处发生碱基变化。结论本实验成功克隆了广西巴马小型猪GHRH成熟肽序列。

Cloning and Sequence Analysis of Growth Hormone Release Hormone (GHRH) Mature Peptide from Guangxi Bama Minipig

Objective To clone and analysis the sequence of GHRH mature peptide gene of Guangxi Bama minipig.Methods The sequence of GHRH exon3 was amplified by a specific pair of primers using Guangxi Bama minipig genome as template.After sequencing the GHRH exon3,the GHRH mature peptide was cloned by over-bang extension PCR protocol and then ligated into sub-clone vector for sequencing and restriction analysis.Results It was showed that the cloned GHRH exon3 and mature peptide gene from Guangxi Bama minipig contained 132 and 105 bp,respectively.Conclusion Sequencing confirmed that the sequence of GHRH exon3 and mature peptide of Guangxi Bama minipig were successfully cloned in the study.