版纳微型猪近交系CATSPER3基因克隆、序列及发育进化分析

Cloning,Sequence and Phylogenetic Analysis of CATSPER3 Gene in Banna Mini-pig Inbred Line

获得版纳微型猪近交系(BMI)CATSPER3基因部分编码区序列,通过生物信息学分析其氨基酸序列和进行不同物种间的同源性比较。以版纳微型猪近交系的公猪睾丸为材料提取RNA,RT-PCR方法扩增CATSPER3基因部分编码区序列,利用在线分析软件进行生物信息学分析。结果显示,扩增出CATSPER3基因部分编码区序列。生物信息学分析表明,推导氨基酸序列,编码蛋白分子量为19.397 3 k D,理论等电点为5.05;在氨基酸组成上,酸性氨基酸(Asp+Glu)有24个、碱性氨基酸(Lys+Arg)有16个,其中以亮氨酸(Leu)占13.3%、缬氨酸(Val)占9.6%、苏氨酸(Thr)占9.0%、苯丙氨酸(Phe)占8.4%、谷氨酸(Glu)占7.2%,天冬氨酸(Asp)占7.2%等含量较高。在核苷酸相似度上与普通猪相似度最高,与山羊核苷酸的相似度较低;分子系统进化树表明与非近交系猪同处于一个分支中,亲缘关系较近,与山羊和绵羊亲缘关系较远。

Our purpose of this work is to obtain the partial coding region sequences of CATSPER3 gene from Banna Mini-pig InbredLine（BMIL）,and to analyze the amino acid sequence and homology comparison among different species by bioinformatics. We used boartestis material of BMIL to extract RNA,RT-PCR to amplify the coding sequence of CATSPER3 gene,and online analysis software to analyzethe bioinformatics. Bioinformatics analysis showed that the protein molecular weight of deduced amino acid sequence was 19.397 3 k D andtheoretical isoelectric point was 5.05. Regarding the composition of amino acid,there were 24 acidic amino acids（Asp and Glu）;and therewere 16 alkaline amino acid（Lys＋Arg）;among them the leucine（Leu）accounted for 13.3%,valine（Val）9.6%,threonine（Thr）9.0%,phenylalanine（Phe）8.4%,glutamate（Glu）7.2%,aspartic acid（Asp）7.2%,and their contents were high. Compared with ordinarypig,the nucleotide similarity was the highest,however,lower with that of goat. Molecular phylogenetic tree showed that inbred pig and noninbred pig clustered in the same branch,i.e.,their genetic relationship was close,while far from goat and sheep.