广西德保猪抵抗素基因的克隆与分析  

Cloning and analysis of Resistin gene in Guangxi Debao pig

作　　者：廖玲玲 李俊[2] 杨厚德 刘庆友[2] 谢炳坤 崔奎青[2] 

机构地区：[1]广西畜牧研究所/广西家畜遗传改良重点实验室,南宁530001 [2]广西大学动物繁殖研究所/亚热带生物资源保护利用国家重点实验室,南宁530005

出　　处：《南方农业学报》2014年第7期1276-1280,共5页Journal of Southern Agriculture

基　　金：国家自然科学基金项目(31360544);广西家畜遗传改良重点实验室开放基金项目(2013GXKFKT-08)

摘　　要：【目的】克隆广西德保猪抵抗素(Resistin,RENT)基因并进行系统生物信息学分析,阐明德保猪脂肪细胞分化与脂肪生成的分子机制。【方法】根据NCBI已公布的猪RENT基因序列(NM_213783.2)设计特异性引物,以从德保猪血液中提取的总RNA为模板,PCR扩增RENT基因序列,T-A克隆至pEASY-T1载体后进行测序分析,并将所得测序结果用BLAST程序、MEGA 5.0软件、EXPASY服务器、SMART程序、SignIP程序、Softberry服务器和DNASTAR软件进行生物信息学分析。【结果】扩增获得468 bp的RENT基因序列片段,包含全长的德保猪RENT基因编码区和部分非编码区序列。多重序列比较分析结果显示,德保猪RENT基因编码区核酸序列与猪、牛、山羊、绵羊、人、兔、猕猴和豚鼠的同源性分别为100.0%、85.5%、85.5%、84.5%、81.2%、80.3%、80.0%和77.3%。德保猪RENT蛋白结构预测结果显示,其理论分子质量11.69 ku,等电点7.82,为弱碱性蛋白;N-末端存在信号肽,定位于胞质外,仅具有1个低复杂度结构域,包含1个α-螺旋、8个β-折叠、7个T-转角及两个无规则卷曲。【结论】RENT基因在德保猪中呈高度保守特征,是其脂肪细胞分化中的重要调控基因。[Objective] Cloning and bioinformatics analysis of Resistin gene in Debao pig were conducted to explore molecular mechanism of adipocyte differentiation and lipogenesis of Debao pig.[Method]With the released pig RENT gene sequence（NM_213783.2） as a template,a pair of primers was designed.The RENT gene was amplified by RTPCR from isolated total RNA of Debao pig blood.The RENT gene was inserted into pEASY-T1 vector by T-A clone method,and was sequenced.And its bioinformatics analysis was performed using BLAST program,MEGA5.0 software,EXPASY server,SMART program,SignIP program,Softberry server and DNAStar program.[Result]A 468bp RENT gene fragment of Debao pig was cloned and sequenced.The sequence multi-aligned results showed that Debao pig RENT gene shared 100.0％,85.5％,85.5％,84.5％,81.2％,80.3％,80.0％ and 77.3％ homology with that of Sus scrofa,Bos taurus,Capra hircus,Ovis aries,Homo sapiens,Oryctolagus cuniculus,Macaca mulatta and Cavia porcellus,respectively.The further protein structure bioinformatics analysis suggested that it was a weakly basic protein with the weight of 11.69 ku and isoelectric point of 7.82.It had an N-terminal signal peptide,localized in the extranuclear with a low complexity domain containing 1α-helices,8 β-folds,7 T-corners and 2 random coils.[Conclusion]Resistin gene fragment in Debao pig is highly conserved in multiple species,and it is of great importance in controlling adipose cell differentiation of Debao pig.

关 键 词：德保猪 RENT基因 克隆 基因功能分析 同源性 蛋白结构预测 

分 类 号：S828.89[农业科学—畜牧学]