CAJ | 학술논문

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.
목적 분리호북정라적미위성DNA서렬,사선다태적미위성DNA위점병분석기특정.방법 응용호북정라기인조DNA적매절편단여생물소표기적(AAT)_(17)、(GA)_(25)、(CCT)_(17)、(CA)_(25)등10개과핵감산탐침잡교,부집、농축、극륭병측서,구건미위성DNA고.도선합괄적미위성DNA위점설계병합성인물,확증정라양본경취병희선알응효전영사선다태성.결과 획득호북정라미위성DNA서렬205조,GenBank주책등기호GU204044～GU204248,기중완정중복서렬74조,점36.10%;비완정중복서렬102조,점49.76%;복합중복서렬29조,점14.15%.설계합성적20대미위성DNA위점인물중,경감정현시13개위점구유다태성.결론 분리건립료호북정라미위성DNA서렬고,위호북정라군체유전、충군소원등상관연구제공료분자표지.
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.