黄胸鼠微卫星分子标记的筛选  

Isolation of microsatellite markers in Rattus tanezumi

作　　者：郭颂[1] 鲁亮[1] 马怀雷[1] 刘起勇[1] 

机构地区：[1]中国疾病预防控制中心传染病预防控制所媒介生物控制室,传染病预防控制国家重点实验室,北京102206

出　　处：《中国媒介生物学及控制杂志》2012年第3期198-201,共4页Chinese Journal of Vector Biology and Control

基　　金：国家科技重大专项课题(2008ZX10004-010)~~

摘　　要：目的研究黄胸鼠不同地理区域种群的遗传差异,用磁珠富集法筛选其微卫星位点。方法将黄胸鼠基因组DNA用限制性内切酶消化后的小片段与接头连接,再用生物素标记的(AC)15重复序列探针与酶切片段杂交,应用链霉素亲和磁珠捕获300～600bp含有微卫星序列的DNA片段,通过载体转化到JM109感受态细胞中,构建富集微卫星序列的小片段插入文库,测序后设计引物并筛选。结果本次实验从约900个转化子中获得304个阳性克隆,对其中140个进行测序,并成功设计黄胸鼠微卫星引物13对。结论筛选出的13对微卫星引物多态性高,能稳定扩增出目标条带,可用于种群遗传研究。Objective To assess the genetic diversity of Rattus tanezumi populations in different geographical regions by screening of microsatellite loci using magnetic beads enrichment. Methods Linkers were ligated with R. tanezumi genomic DNA fragments obtained by digestion by Hae III and Rsa I, and the effect of ligation reaction was detected by PCR. Then the fragments were hybridized with biotinylated （AC）15 probe, and the 300-600 bp fragments containing microsatellite markers were captured by magnetic beads, and then the PCR products of the captured fragments were transformed into JM109 competent ceils, with microsatellite primers designed after being sequenced. Results A total of 304 positive colonies were obtained from about 900 transformants and 140 of them were sequenced with 13 pairs of microsatellite primers successfully designed for R. tanezumi. Conclusion The acquired microsatellite primers are polymorphic and can amplify bands stably, being fit for the research of population genetics of R. tanezumi.

关 键 词：黄胸鼠 磁珠富集 微卫星 

分 类 号：S443[农业科学—农业昆虫与害虫防治]