雷氏按蚊多态微卫星DNA位点的筛选和特征

本研究对雷氏按蚊的微卫星DNA序列进行了分离,并筛选了其中具有多态性的微卫星位点.应用雷氏按蚊基因组DNA的酶切片段与生物素标记的寡核苷酸探针杂交,亲合素富集和超滤离心浓缩目的片段,扩增放大后克隆并测序,获得雷氏按蚊微卫星DNA序列.在其中挑选合适的微卫星位点,建立扩增体系.应用雷氏按蚊现场样本对不同的微卫星DNA进行扩增,聚丙烯酰胺凝胶电泳筛选具有多态性的位点.本研究结果获得了雷氏按蚊微卫星DNA序列58条,GenBank注册登记号为EF620299～EF620356.其中,完整序列41条,占70.7%;非完整序列7条,占12.1%,复合序列9条,占15.7%;仅1条为非典型序列.电泳结果显示其中14个微卫星位点中有11个具有多态性.

ISOLATION AND CHARACTERIZATION OF POLYMORPHIC MICROSATELLITE MARKERS OF ANOPHELES LESTERI (DIPTERA: CULICIDAE)

Microsatellite DNA sequences of Anopheles lesteri were isolated,and the polymorphic microsatellite loci were characterized. Digested genomic fragments were hybrizied with biotinylated oligonucleotide probes. The target fragments moleculars were captured with Vectrex Avidin D and enriched by centrifuged with ultra-4-column ultra-filtrate. Then these fragments containing microsatellite repeats were amplified,cloned and sequenced. The suitable microsatellite loci were chosen and established with the PCR amplification assay. The polymorphism screening was used by PAGE gel electrophoresis with wild individuals of An. lesteri. A total of 58 microsatellite DNA sequences were obtained in this study (GenBank Acession numbers: EF620299-EF620356). The percentage of perfect microsatellite DNA sequence was 70.7% (41/58),with imperfect sequence as 12.1% (7/58) and compound sequence as 15.7% (9/58). Only one was non-typical microsatellite sequence. 11 of 14 microsatellite loci were found.