版纳微型猪近交系AO血型A基因的克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of AO Blood Group A Gene in Banna Mini-pig Inbred Line

作　　者：潘伟荣[1,2] 查星琴 成文敏[1,2] 卿玉波 肖晶[2] 魏红江 曾养志[2] 霍金龙 PAN Weirong;ZHA Xingqin;CHENG Wenmin;QING Yubo;XIAO Jing;WEI Hongjiang;ZENG Yangzhi;HUO Jinlong(Animal Science and Technology College, Yunnan Agricultural University, Ktmming 650201, China;Key Laboratory of Banna Mini-pig Inbred Line of Yunnan Province, Kunming 650201, China)

机构地区：[1]云南农业大学动物科学技术学院,云南昆明650201 [2]云南省版纳微型猪近交系重点实验室,云南昆明650201

出　　处：《云南农业大学学报（自然科学版）》2018年第1期56-62,共7页Journal of Yunnan Agricultural University:Natural Science

基　　金：云南省教育厅科学研究基金项目(2014Z077)

摘　　要：【目的】扩增版纳微型猪近交系AO血型A基因编码区全长序列,并对序列进行生物信息学分析。【方法】提取版纳微型猪近交系猪组织的RNA,反转录成为cDNA,利用RT-PCR方法扩增AO血型A基因编码区全长序列,将纯化的片段与pMD18-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】版纳微型猪近交系AO血型A基因编码区全长1 095 bp,编码364个氨基酸,氨基酸序列与GenBank上登录的猪AO基因氨基酸序列相似性为99.5%。【结论】版纳微型猪近交系的AO血型A基因除了与黑猩猩、人和小鼠亲缘关系较远外,与牦牛、藏羚羊、水牛、小须鲸、绵羊、羊驼、野骆驼、山羊的关系都较为接近,其中与山羊的亲缘关系最为接近。[Purpose]To amplify the encoding full-length sequence of AO blood group A gene from Banna mini-pig inbred line and analysis the sequence by bioinformatics software.[Method]The tissues RNA from Banna mini-pig inbred line was extracted, and reversely transcribed into cDNA. The gene coding region sequence of AO blood group A gene was amplified through RT-PCR, and the purified product of PCR was ligated with a pMD18-T vector and transferred into the bacterium DH5α 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 region of AO blood group A gene was 1 095 bp, and coded 364 amino acids. The amino acid homology analysis showed that the homology rate of amino acid sequence was 99.5% with that of other species in GenBank.[Conclusions]Molecular phylogeny evolution indicated that it had a close relation with yak, Tibetan antelope, buffalo,minke whale, sheep, alpaca, wild camel, goat except chimpanzee, human and mouse, and the most closely related with goat.

关 键 词：版纳微型猪近交系 AO血型A基因 克隆测序 生物信息学 

分 类 号：S828.2[农业科学—畜牧学]