大骨鸡胚胎肌生成抑制素基因(MSTN)cDNA的克隆、测序及原核表达  

Gene cloning and prokaryotic expression of MSTN cDNA in big bone chicken embryo

作　　者：胡兰[1] 李桂伶[1] 吴丹[1] 郝文博[1] 杨晓宇[1] 董顺义[1] 

机构地区：[1]沈阳农业大学畜牧兽医学院,辽宁沈阳110161

出　　处：《中国兽医学报》2009年第9期1170-1173,共4页Chinese Journal of Veterinary Science

基　　金：辽宁省自然科学基金资助项目(20050302);辽宁省教育厅科研基金资助项目(2007304)

摘　　要：提取大骨鸡胚胎腿肌RNA，对肌生成抑制素基因（MSTN）RT—PCR扩增、TA克隆和测序。结果表明，大骨鸡胚胎腿肌中存在2种不同大小的MSTN cDNA，一种MSTN cDNA由1128bp组成，我们将该序列称为chMSTN-1，与已报道的鸡MSTN cDNA（gi：38349516）序列同源性为100％；另外一种MSTN cDNA由985bp组成，与chMSTN-1相比，缺失了374～517处的143个碱基，并且在51，234，324bp处也发生碱基变化，这可能是一种新的MSTN cDNA，我们将该序列命名为chMSTN-2。另外，分别对上述2种MSTN cDNA进行双酶切、亚克隆，成功构建重组表达载体pGEX—KG-chMSTN-1与pGEX-KG-chMSTN-2，转染大肠杆菌并诱导表达。检测结果表明，chMSTN-1和chMSTN-2均能在大肠杆菌中表达，重组蛋白以包涵体形式存在，大小分别约为66000和55000。RT-PCR was used to study the expression of myostatin. The results showed there were two myostatin gene transcriptons of varied sizes in leg muscle of big bone chicken embryo by cloning and sequencing. One myostatin mRNA was composed of 1 128 bp,and named chMSTN-1, which held 99% homology with the chicken myostatin gene mRNA reported （gi：38349516）;the other myostatin gene eDNA was consisted of 985 bp with a homology of 87% with chMSTN-1 ,and 143 bases from 374 to 517 were deleted as compared with chMSTN-1 ,and the bases were changed at sites of 51,234 and 324 bp. It May be a new myostatin gene eDNA,and named chMSTN-2. TA cloning with two myostatin cDNAs of big bone chicken and pMD-18 T vector,then the eDNA to prokaryotic expression vector pGEX-KG and expression in E. coli. It was demonstrated in the results that,chMSTN-1 could express 66 000 protein in E. coli,while chMSTN-2 could express 55 000 protein.

关 键 词：肌生成抑制基因cDNA 大骨鸡 原核表达 

分 类 号：S852.23[农业科学—基础兽医学]