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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。
Objective To isolate the microsatellite DNA sequence from Hubei snails and screen the polymorphic microsatellite DNA sites and analyze their characteristics. Methods A total of 10 oligonucleotide probes, such as biotin-labeled (AAT) 17, (GA) 25, (CCT) 17 and (CA) Cloned and sequenced to construct a microsatellite DNA library. Select the appropriate microsatellite DNA site design and synthesis of primers, amplified snail samples by polyacrylamide gel electrophoresis screening polymorphisms. Results There were 205 microsatellite DNA sequences of Oncomelania hupensis in GenBank with GU204044 ~ GU204248, including 74 complete repeats (36.10%), 102 non-complete repeats (49.76%), 29 composite repeats (14.15% . Of the 20 pairs of microsatellite DNA loci designed and synthesized, 13 loci were identified as being polymorphic. Conclusion The microsatellite DNA sequence library of Oncomelania hupensis was established and isolated, which provided the molecular markers for the study of population genetic and population tracing of Oncomelania hupensis in Hubei Province.