黄胸鼠微卫星分子标记的筛选

中国媒介生物学及控制杂志 ›› 2012, Vol. 23 ›› Issue (3): 198-201.

黄胸鼠微卫星分子标记的筛选

郭颂, 鲁亮, 马怀雷, 刘起勇

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

收稿日期:2011-12-30 出版日期:2012-06-20 发布日期:2012-06-20
通讯作者: 鲁亮,Email: luliang@icdc.cn
基金资助:
国家科技重大专项课题(2008ZX10004-010)

Isolation of microsatellite markers in Rattus tanezumi

GUO Song, LU Liang, MA Huai-lei, LIU Qi-yong

State Key Laboratory for Infectious Diseases Prevention and Control, Department of Vector Biology and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China

Received:2011-12-30 Online:2012-06-20 Published:2012-06-20
Supported by:
Supported by the Major National Science and Technology Projects of China(No. 2008ZX10004-010)

摘要/Abstract

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300～600 bp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。

关键词: 黄胸鼠, 磁珠富集, 微卫星

Abstract: 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 HaeⅢ and RsaⅠ, 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 cells, 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.

Key words: Rattus tanezumi, Magnetic beads enrichment, Microsatellite

中图分类号:

S443

郭颂, 鲁亮, 马怀雷, 刘起勇. 黄胸鼠微卫星分子标记的筛选[J]. 中国媒介生物学及控制杂志, 2012, 23(3): 198-201.

GUO Song, LU Liang, MA Huai-lei, LIU Qi-yong. Isolation of microsatellite markers in Rattus tanezumi[J]. Chines Journal of Vector Biology and Control, 2012, 23(3): 198-201.

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