版纳微型猪近交系SRY基因编码区序列克隆及生物信息学分析

Cloning and bioinformatics analysis of coding region of the SRY gene in Banna mini-pig inbred line(BMI)

目的获得版纳微型猪近交系(BMI)SRY基因编码区序列并进行分子系统进化研究。方法以看家基因GAPDH为内参,对BMI猪的SRY基因编码区序列进行PCR扩增、克隆和序列分析,并应用Lasergene、Bi-oEdit、ClustalX、MEGA等生物信息学软件同鲸鱼、海豚、鹿、绵羊、牛、海豹、马、海象、熊猫、人、驴、熊、猫、虎和美洲豹等15个物种相应SRY编码区核苷酸序列和氨基酸序列进行了比对分析,在此基础上采用NJ和ME法对其编码区氨基酸序列构建了分子系统进化树。结果 BMI猪SRY基因编码区序列长711 bp,编码236个氨基酸,GenBank登录号为GU991615。BMI猪与鲸鱼、海豚、鹿、绵羊、牛、海豹、马、海象、熊猫、人、驴、熊、猫、虎和美洲豹的SRY基因编码区核苷酸序列相似性分别为83.7%、82.8%、78.4%、78.0%、76.9%、73.3%、73.1%、73.0%、72.9%、72.7%、72.7%、72.2%、71.6%、71.3%、70.8%,相应的氨基酸序列相似性分别为72.8%、54.5%、67.3%、64.5%、61.5%、61.9%、59.5%、61.4%、62.0%、59.1%、59.0%、62.0%、59.6%、59.6%、59.2%。结论 BMI猪同鲸鱼等15个物种的SRY基因编码区核苷酸和相应氨基酸序列具有较高的保守性。NJ和ME方法聚类构建的分子系统进化树表明,BMI猪与牛、绵羊、鹿聚为一个分支,符合分类学地位,它们分别为偶蹄目下的猪科、牛科和鹿科动物。

Objective To obtain the coding region sequence the SRY gene of Banna mini-pig inbred line（BMI） and construct the molecular phylogenetic tree.Methods The coding region of the SRY gene of BMI was cloned and sequenced by using housekeeping gene GAPDH as internal control,then based on the nucleotide and amino acid sequence to compare with some species（whale,dolphin,deer,sheep,cattle,seal,horse,walrus,panda,human,ass,bear,cat,tiger and leopard） by Lasergene,BioEdit,ClustalX,Mega bio-softs,and also constructed the molecular phylogenetic tree on coding amino acids region of SRY gene with method of NJ and ME.Results The sequence length of coding region of the BMI SRY gene was 711 bp,with GenBank accession number GU991615,and encoding 236 amino acid.The similarity rates of nucleotide sequences of the coding region of SRY gene between the BMI and whale was（83.7%）,dolphin（82.8%）,deer（78.4%）,sheep（78.0%）,cattle（76.9%）,seal（73.3%）,horse（73.1%）,walrus（73.0%）,panda（72.9%）,human（72.7%）,ass（72.7%）,bear（72.2%）,cat（71.6%）,tiger（71.3%） and leopard（70.8%）;while the similarity rates of amino acid sequences were 72.8%,54.5%,67.3%,64.5%,61.5%,61.9%,59.5%,61.4%,62.0%,59.1%,59.0%,62.0%,59.6%,59.6% and 59.2%,respectively.Conclusions The nucleotide sequence of the coding region and accordingly amino acid sequences of SRY gene of BMI and whale,dolphin,deer,sheep,cattle,seal,horse,walrus,panda,human,ass,bear,cat,tiger and leopard are quite conservative.NJ and ME molecular phylogenetic tree indicate that the taxonomic placement of BMI is clustered together with cattle,sheep and deer,which are the suidae,bovidae,cervidae,respectively,and all of them are artiodactyla animals.