广西巴马小型猪肥胖基因(leptin)成熟肽序列的cDNA克隆、序列分析  

Molecular cloning and sequencing of leptin gene mature peptides cDNA from Guangxi Bama mini-pig

作　　者：兰干球[1] 李芳芳[1] 黄雄军[1] 郭亚芬[1] 王爱德[1] 李柏[1] 

机构地区：[1]广西大学动物科学技术学院,广西南宁530005

出　　处：《广西农业生物科学》2008年第3期183-186,共4页Journal of Guangxi Agricultural and Biological Science

基　　金：广西科技攻关项目(桂科攻0428011-1)

摘　　要：以广西巴马小型猪的脂肪组织总RNA为模板,通过RT-PCR扩增出巴马小型猪的leptin基因成熟肽序列,经纯化回收后连接到pMD18-T载体上,经鉴定后测序。研究结果表明,本试验克隆到leptin基因成熟肽cDNA序列,其长度为441 bp,与GenBank中报道的猪leptin成熟肽cDNA序列同源性高达99.06%,与奶牛、家鼠、马等物种的成熟肽cDNA序列间同源性可达到84.5%以上,证明leptin基因具有较高的保守性。同时发现,与普通猪相比,巴马小型猪leptin基因成熟肽cDNA序列有四处存在碱基突变,均为无义突变。The total RNA was purified from Guangxi Bama Mini-pig subcutaneous adipose tissue. A fragment with 441 bp was amplified by the Reverse Transcription Polymerase Chain Reaction （RT- PCR） and then was ligated into pMD18-T vector after purification. The recombinant was verified by the method of PCR, restrictive cleavage and sequencing, respectively. The result showed that the Bama Mini-pig leptin gene mature peptide cDNA was successfully cloned in this study. The homologous analysis indicated that the homologous rate was 99. 06 % compared with the porcine leptin gene reported in GenBank and this gene has high conservativeness among mammalian animals （〉84. 5 %）. Four nonsence mutations were detected in leptin gene mature peptide cDNA sequence of Bama Mini-pig.

关 键 词：广西巴马小型猪 LEPTIN基因 克隆 

分 类 号：Q781[生物学—分子生物学]