崂山奶山羊Myostatin基因真核表达载体的构建及其在成纤维细胞中的表达研究

Construction of Eukaryotic Expression Vector of Laoshan Dairy Goat Myostatin and its Expression in the Fibroblast Cells

试验克隆了崂山奶山羊Myostatin基因序列,构建了其真核表达载体,并验证了其在成纤维细胞中的表达。本研究通过从崂山奶山羊肌肉组织中提取RNA,反转录后采用巢式PCR方法扩增出Myostatin基因序列,构建真核表达载体,通过转染成纤维细胞,采用RT-PCR方法验证其表达。结果表明,克隆出崂山奶山羊Myostatin基因的全长cDNA序列,大小为1128bp,GenBank登录号:GU377303.1;构建pcDNA-MSTN真核表达载体,转染成纤维细胞48h后通过RT-PCR检测,结果显示,Myostatin表达量显著增加,表明成功构建pcDNA-MSTN真核表达载体,为进一步研究Myostatin的生物学功能及转基因羊培育奠定基础。

This study aimed to clone cDNA of Myostatin of Laoshan dairy goat,and to construct Myostatin eukaryotic expression vector,analyzing its expression in the fibroblast cells.RNA was extracted from skeletal muscle and then reverse transcripted into cDNA.Myostatin was amplified by nested PCR.Myostatin was cloned into pcDNA3.1 to synthesize an eukaryotic expression vector,pcDNA-MSTN and its expression was detected by RT-PCR after transfected into the fibroblast cells.The results showed that the sequence of the full length cDNA of Myostatin gene of Laoshan dairy goat was similar with other species of goats and GenBank accession number was GU377303.1.The expression of Myostatin was significantly increased as detected by RT-PCR in fibroblast cells transfected with the expression vector.The study provided a basis for further study of the biological function of Myostatin and transgenic goat.