摘要
目的探讨湖北、广东、辽宁和韩国4地华支睾吸虫的基因差异,为华支睾吸虫种下分型提供依据。方法取湖北、广东两地的华支睾吸虫,提取基因组DNA,PCR特异性扩增18S rDNA V4区并测序;登陆GenBank,检索中国辽宁（AF217100）和韩国（AF408144）的华支睾吸虫18S rDNA的登录序列,应用系统发生学分析法对所有序列进行排列比较,分析4地华支睾吸虫的基因差异,并构建系统发生树,分析其亲缘关系。结果获得的湖北和广东两地华支睾吸虫18S rDNA V4区的碱基数分别为392 bp和440 bp。通过对18S rDNA V4区基因序列的比较与进化树的构建,证实4个地域株华支睾吸虫的18S rDNA V4区具有较高的同源性（98%~100%）,彼此间遗传距离较小（0~0.013）;在系统发生树中,湖北株和广东株华支睾吸虫形成一个支系,韩国株和辽宁株华支睾吸虫形成另外一个支系。结论以18S rDNA V4区为分子标记的DNA序列分析表明,湖北、广东、辽宁和韩国4地华支睾吸虫存在基因差异,但亲缘关系较近,即起源于共同的祖先。
Objective To study the genetic divergence of Clonorchis sinensis in Hubei,Guangzhou,Shengyang,and South Korea and lay the groundwork for C.sinensis classification.Methods The genomic DNA of C.sinensis from Hubei and Guangdong was extracted and the 18S rDNA V4 region of these trematodes was specifically amplified.The PCR products were sequenced.The 18S rDNA V4 region of C.sinensis from Shengyang and South Korea was found in GenBank and these sequences were aligned and compared using phylogenetic analysis.The differences in these sequences were analyzed.The 18S rDNA phylogenetic tree was constructed and the kinship between these 4 strains was analyzed.Results The 18S rDNA V4 region of C.sinensis ranged from 392 bp to 440 bp in size.Among the 4 strains,the homology of the 18S rDNA V4 region was 98%-100% and the genetic distance ranged from 0 to 0.013.In the phylogenetic tree,the strains from Hubei and Guangdong form one branch and the strains from Shengyang and South Korea form another branch.Conclusion The 18S rDNA V4 region is a useful way to study the phylogeny of C.sinensis.There was genetic variance in the 4 strains of C.sinensis studied,but the 4 strains of C.sinensis shared a close kinship,indicating that they belong to the same species and originated from the same ancestor.
出处
《中国病原生物学杂志》
CSCD
2011年第6期443-445,I0002,共4页
Journal of Pathogen Biology
