【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。更多还原

【Abstract】 [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.更多还原