摘要
【目的】克隆内蒙古绒山羊SRY基因的HMG-box区,并对其序列特征进行分析。【方法】参考Gen-Bank中山羊SRY基因序列设计1对引物,采用PCR法扩增内蒙古绒山羊雄性个体的SRY基因HMG-box区全部序列,克隆到pMD18-T载体中,转化DH5α菌,蓝白斑筛选阳性克隆,并进行测序。将测序结果与GenBank中部分偶蹄目动物SRY基因HMG-box的序列进行同源性比较,构建系统进化树。【结果】内蒙古绒山羊SRY基因HMG-box长度为231 bp,编码77个氨基酸;内蒙古绒山羊SRY基因HMG-box与GenBank中山羊、绵羊、猪、麝香鹿、梅花鹿、牛、水牛和牦牛等偶蹄目动物的核苷酸同源性分别为100.0%,99.1%,86.1%,96.1%,96.1%,97.4%,95.6%和96.9%,推导的氨基酸序列同源性分别为100.0%,100.0%,83.1%,94.8%,94.8%,94.8%,92.2%和93.5%。基于SRY基因HMG-box核苷酸序列构建的物种间系统进化结果,基本上与这些偶蹄目动物传统的系统分类结果一致。【结论】SRY基因HMG-box区在物种进化中高度保守,有望作为构建系统发育树的候选基因。
[Objective] This study aim to clone HMG-box of SRY gene in Inner Mongolia cashmere goat,and analyzed its sequence characteristics. [Method] From male Inner Mongolia cashmere goat genomic DNA,the fragment of SRY gene containing entire HMG-box region was amplified by PCR with a pair of designed primers based on the sequence of goat SRY gene sequence from GenBank. The amplified fragment was cloned to the pMD18-T vector,and transformed to the bacterium DH5α. The identified positive clone with amplified fragment was sequenced. The sequences homology of SRY gene HMG-box region of some artiodactylas from GenBank was compared with the Inner Mongolia cashmere goat, and based on the sequences of HMG-box region, the phylogenetic tree was constructed by UPGMA method. [Result] The HMG-box region of Inner Mongolia cashmere goat SRY gene was 231 bp,encoding 77 amino acid residues. The corresponding sequences in GenBank of goat, sheep, pig, musk deer, sika deer, cattle, water buffalo and yak,had homology of 100.0%,99.1% ,86.1%,96.1%,96.1%,97.4%,95.6% and 96.9% in nucleotide respectively with the Inner Mongolia cashmere goat, and had homology of 100. 0%, 100. 0%,83. 1%, 94.8%,94.8%,94.8%,92.2% and 93. 5% in induced amino acid respectively with the Inner Mongolia cashmere goat. The clustering based on the sequences of SRY gene HMG-box region among species was generally in agreement with the classic taxonomic relationship. [Conclusion] The HMG-box of SRY gene is rather conservative in evolution of species,and might be useful in the construction of phylogenetic tree among species.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2009年第7期44-50,共7页
Journal of Northwest A&F University(Natural Science Edition)
基金
辽宁省教育厅科学研究基金计划项目(2005358)
