西藏小型猪MSTN基因分子克隆及序列分析  

Cloning and sequence analysis of Tibet mini-pig's MSTN gene

作　　者：吴清洪[1] 岳敏[1] 田雨光[1,2] 王玉珏[1] 徐名衬[1] 顾为望[1,2] 

机构地区：[1]南方医科大学实验动物中心比较医学研究所,广州510515 [2]东莞松山湖明珠实验动物科技有限公司,广东东莞523808

出　　处：《中国比较医学杂志》2015年第3期18-22,共5页Chinese Journal of Comparative Medicine

基　　金：广东省科技计划项目(2012B011000004;2012B010300001);广东省科技厅科技基础条件建设项目(2010B060500001);国家科技部国际合作项目(2011DFA33290)

摘　　要：目的扩增西藏小型猪的肌肉生长抑制素(MSTN)基因编码区全长序列,并对序列进行生物信息学分析。方法提取西藏小型猪肝脏组织的RNA,反转录成为c DNA,利用RT-PCR方法扩增MSTN基因编码区全长序列,将纯化的片段与pMD19-T载体连接,转化大肠杆菌DH5α,筛选阳性克隆并测定其序列,利用生物信息学方法分析其序列特征,并与其他12种物种构建系统进化树。结果西藏小型猪MSTN基因编码区全长1128bp,编码375个氨基酸,氨基酸序列同源性分析表明,与版纳微型猪MSTN氨基酸序列相似性最高,为99%。结论 MSTN基因西藏小型猪除了与鸡和斑马鱼亲缘关系较远外,与人、犬、版纳微型猪、绵羊、山羊、牛、马、黑猩猩、大鼠、小鼠的关系都较为接近,其中与版纳微型猪的亲缘关系最为接近。Objective To amplify the encoding full-length sequence of Tibet mini-pigs myostatin （MSTN） gene and analysis the sequence by bioinformatics software. Method The RNA of liver tissues from Tibet mini-pig was extracted, and reversely transcribed into eDNA. The gene coding region sequence of myostatin gene was amplified through RT-PCR, and then the purified product of PCR was ligated with a pMD19-T and transferred into the bacterium DH5a for replication. The positive clones were screened and sequenced. The sequence characters were analyzed by using bioinformatics method and phylogeny evolution tree was constructed with other twelve species. Results The coding redion of MSTN gene was 1128bp, and coded 375 amino acids. The amino acid homology analysis showed that the homology rate of amino acid sequence was 99%. Conclusions Molecular phylogeny evolution indicated that it had a close relation with human, dog, banna minipig, sheep, goat, cattle, horse, chimpanzee, rat, mouse except chick and zebrafish, and the most closely related with banna minipig.

关 键 词：西藏小型猪 MSTN基因 克隆测序 

分 类 号：R332[医药卫生—人体生理学]