湖北钉螺多态微卫星DNA位点筛选和特征初步分析

目的分离湖北钉螺的微卫星DNA序列,筛选多态的微卫星DNA位点并分析其特征。方法应用湖北钉螺基因组DNA的酶切片段与生物素标记的(AAT)17、(GA)25、(CCT)17、(CA)25等10个寡核苷酸探针杂交,富集、浓缩、克隆并测序,构建微卫星DNA库。挑选合适的微卫星DNA位点设计并合成引物,扩增钉螺样本经聚丙烯酰胺凝胶电泳筛选多态性。结果获得湖北钉螺微卫星DNA序列205条,GenBank注册登记号GU204044～GU204248,其中完整重复序列74条,占36.10%;非完整重复序列102条,占49.76%;复合重复序列29条,占14.15%。设计合成的20对微卫星DNA位点引物中,经鉴定显示13个位点具有多态性。结论分离建立了湖北钉螺微卫星DNA序列库,为湖北钉螺群体遗传、种群溯源等相关研究提供了分子标志。

Isolation and characterization of polymorphic microsatellite markers of Oncomelania hupensis

Objective To isolate the microsatellite DNA sequences of Oncomelania hupensis and analyze the polymorphic microsatellite loci.Methods The digested genomic fragments were hybridized with biotinylated oligonucleotide probes.The target fragments moleculars were captured and enriched.Then these fragments were cloned and sequenced.The suitable microsatellite loci were chosen and the polymorphism was screened by PAGE gel electrophoresis.Results A total of 205 microsateilite DNA sequences were obtained (GenBank accession numbers :GU204044～GU204248).The percentage of perfect microsatellite DNA sequence was 36.10% (74/205),with imperfect sequence as 49.76% (102/205) and compound sequence as 14.15% (29/205).Twenty typical microsatellite sequences were selected to design amplifying primers,and 13 microsatellite loci were found to be polymorphism.Conclusion A total of 205 microsatellite DNA sequences of Oncomelania hupensis are isolated and first reported,which will be useful for population genetic and mapping studies of Oncomelania hupensis.