牦牛MSMB基因的克隆及其生物信息学分析  

Cloning and Bioinformatics Analysis of Yak MSMB Gene

作　　者：刘宇 何向东 夏忆 陈莹[1] 字向东[1] Liu Yu;He Xiangdong;Xia Yi;Chen Ying;Zi Xiangdong(The Key Laboratory of Animal Science of State Ethnic Affairs Commission,Southwest Minzu University,Chengdu 610041,China)

机构地区：[1]西南民族大学动物科学国家民委重点实验室

出　　处：《中国草食动物科学》2019年第5期1-5,22,共6页China Herbivore Science

基　　金：国家重点研发计划项目(2018YFD0502303);教育部卓越农林人才教育培养计划

摘　　要：微精原蛋白β(MSMB)对动物的繁殖功能具有重要的调控作用。采集成年健康母牦牛的卵巢组织,通过RT-PCR技术克隆牦牛的MSMB基因,并进行了生物信息学分析。结果表明:牦牛MSMB基因CDS区长117 bp,编码38个氨基酸;MSMB蛋白分子式C194H308N50O59S4,理论等电点7.69,分子质量4.4 ku,脂肪系数66.58,不稳定系数21.23,属于稳定蛋白质,半衰期30 h,不具有跨膜结构和信号肽,是亲水性蛋白;与黄牛MSMB基因对比发现,牦牛MSMB基因存在1处碱基突变;牦牛与黄牛和绵羊CDS区核苷酸同源性均在80%以上,说明MSMB基因在进化中保守;MEGA 6.0和Clustal X软件分析结果表明,牦牛与黄牛首先聚为一小类,其次与绵羊、小鼠和大鼠聚为一类,最终与猪、鸡和鱼聚为一类,符合物种进化规律,进化树与同源性比对结果相符;二级结构α螺旋、延长链、β-转角和无规卷曲,分别占13.16%、44.74%、13.16%、28.95%,二级结构与三级结构相符;预测MSMB蛋白其结构域是PSP94超家族成员。该实验结果为进一步研究牦牛MSMB基因功能提供了一定理论依据。Microseminoprotein-β(MSMB)plays a key regulatory role in animal reproductive function.In the present study,the ovary tissues were collected from healthy female yak.The yak MSMB gene was cloned by RT-PCR technology and analyzed by bioinformatics method.The results showed that the CDS region of yak MSMB gene is 117 bp and encoding a 38 amino acid protein.The formula of MSMB protein was C194H308N50O59S4 and its theoretical isoelectric point,molecular weight,and the fat coefficient were 7.69,4.4 ku and 66.58,respectively.MSMB protein was a stable protein with 30 h of half-life and has no transmembrane structure and no signal peptide,indicating a hydrophilic protein.The nucleotide homology of CDS region of yak MSMB compared to cattle and sheep was over 80%,indicating that MSMB gene was conserved in evolution course.Analysis by MEGA 6.0 and Clustal X showed that yaks and cattle were firstly grouped into a small group,then into sheep,mouse and rats,and finally into pigs,chickens and fish.The results of phylogenetic tree and homology were consistent.The secondary structureα-helix,extended strand,β-turn and random coil of MSMB protein accounted for 13.16%,44.74%,13.16%and 28.95%,respectively.The predicted domain of MSMB protein belonged to PSP94 superfamily.This experiment provides a theoretical basis for further study of yak MSMB gene function.

关 键 词：牦牛 MSMB基因 基因克隆 生物信息学分析 

分 类 号：S823.85[农业科学—畜牧学]