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目的研究湖南省日本血吸虫不同地域自然隔离群线粒体ATP合成酶F0亚单位6基因(atp6)部分序列的遗传多态性,为湖南不同地域自然隔离群日本血吸虫的遗传特性研究提供实验依据。方法采用试剂盒抽提基因组总DNA,以特异性引物对线粒体atp6基因进行PCR扩增,通过单链构象多态性技术(SSCP)筛选出差异带型并进行测序,用DNAStar 5.0及Mega 4.0软件进行比对分析。结果湖南省5个流行区的日本血吸虫PCR扩增后获得了483 bpatp6部分序列,检测出17个变异位点,变异率为3.52%,雌雄虫之间的差异为0.0%～1.3%,不同地理来源虫株间的差异率为0.0%～1.5%。聚类分析结果表明:湖南不同地域自然隔离群日本血吸虫的种系发育不能按照其来源归为相应的系统发育进化树。结论湖南不同地域自然隔离群日本血吸虫的atp6基因存在明显的个体差异,是否由于生存环境的选择压力造成了同一地理来源日本血吸虫个体之间atp6基因的遗传差异,其原因尚有待于进一步研究。
Objective To study the genetic polymorphisms of the atp6 part of mitochondrial ATP synthase (F0) subunit 6 gene in different geographical isolates of Hunan Schistosoma japonicum and provide experimental evidence for the genetic characterization of Schistosoma japonicum in different geographical isolates in Hunan Province. Methods Genomic DNA was extracted by kit and PCR amplification of mitochondrial atp6 gene was carried out with specific primers. The different bands were screened by single strand conformation polymorphism (SSCP) and sequenced. DNAStar 5.0 and Mega 4.0 software For comparison analysis. Results The 483 bpatp6 partial sequence was obtained after PCR amplification of S. japonicum in five endemic areas of Hunan Province. Seventeen variation sites were detected, with a variation rate of 3.52% and a difference of 0.0% -1.3% between male and female. The difference between the source strains was 0.0% ~ 1.5%. Cluster analysis showed that the phylogeny of Schistosoma japonicum, a natural isolate in different regions of Hunan Province, can not be classified as phylogenetic tree according to its origin. Conclusions There are obvious individual differences in the atp6 gene of Schistosoma japonicum in natural isolates from different regions of Hunan. Whether the genetic variation of atp6 gene between individuals of the same geographical origin, Schistosoma japonicum, is caused by the selection pressure of living environment remains to be further studied.