摘要
【目的】对陆川猪肌肉生长抑制素(MSTN)基因进行克隆及序列分析,为后期开展MSTN基因表达与陆川猪肌肉生长发育及脂肪沉积的相关性研究奠定基础。【方法】根据GenBank中已发表的猪MSTN基因序列(NM_214435)设计引物,以陆川猪背最长肌组织总RNA为模板,RT-PCR扩增MSTN基因cDNA序列,扩增片段经纯化、克隆、鉴定和测序分析,并应用生物软件进行蛋白质二级结构预测。【结果】克隆得到陆川猪MSTN基因编码区1128 bp,与GenBank已公布的约克夏、汉普夏、杜洛克、梅山猪、藏猪、广西巴马小型猪的基因同源性均在99.8%以上。陆川猪MSTN基因第294位点存在特有的碱基G,但未引起氨基酸改变。蛋白质二级结构预测结果表明,陆川猪的MSTN成熟肽包含有α螺旋、β转角、延长线和无规卷曲4种二级结构元件。【结论】猪MSTN基因高度保守,可作为研究陆川猪肌肉生长发育和脂肪沉积的候选基因之一。
[Objective ]MSTN gene in Luchuan pig was cloned and its sequence was analyzed to provide references for fur- ther research on MSTN gene expression, muscle growth and development and fat deposition of Luchuan pig. [Method]Accord- ing to the pig MSTN gene sequence published in GenBank, a pair of primers was designed to amplify the MSTN gene eDNA of Luchuan pig using RT-PCR. Then the obtained fragments were purified, cloned, indentified and sequenced. The protein sec- ondary structure of MSTN was predicted and analyzed by biological software. [Result]The length of the coding region for the cloned MSTNgene in Luchuan pig was 1128 bp, and the sequence homologies of MSTNgene with the sequence of Yorkshire, Hampshire, Duroc, Meishan, Tibetan and Guangxi Bama minipig were more than 99.8%. In addition, specific base point G existed in the 294~ site, but the mutation did not cause change in amino acid. The results of protein secondary structure pre- diction showed that Luchuan pig MSTN mature peptide contained four secondary structure elements, which were ot helix, 13 comer, extension cords and random coil. [Conclusion]MSTN was highly conservative and can be used as one of the candidate genes for researching muscle growth and development and fat deposition of Luchuan pig.
出处
《南方农业学报》
CAS
CSCD
北大核心
2014年第4期659-663,共5页
Journal of Southern Agriculture
基金
广西大学科研基金项目(XBZ130087)
