Screening of Small-insert Genomic DNA Libraries for Brandt’s voles by PCR

doi:10.11733/j.issn.1007-0435.2007.03.016

›› 2007, Vol. 15 ›› Issue (3): 278-282.DOI: 10.11733/j.issn.1007-0435.2007.03.016

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Screening of Small-insert Genomic DNA Libraries for Brandt’s voles by PCR

WANG Deng, SHI Da-zhao

China Agricultural University, Beijing 100094, China

Received:2006-11-21 Revised:2007-01-26 Online:2007-06-15 Published:2007-06-15

PCR方法筛选布氏田鼠小片断插入微卫星文库

王登, 施大钊

中国农业大学, 北京, 100094

通讯作者: 施大钊,E-mail:s hidazhao@cau.edu.cn
作者简介:王登(1976- ),男,博士研究生,研究方向为啮齿动物种群遗传,E-mail:w.deng2001@163.com
基金资助:
国家自然科学基金(30571229);农业部专项资金(2005BA529A05)

Abstract

Abstract: In order to assess the genetic diversity of the different geographical populations of Brandt’s voles(Lasiopodomys brandtii),we isolated the polymorphic microsatellite loci and constructed small-insert genomic DNA libraries for Brandt’s voles.Genome DNA was digested with Sau3AI to produce 100-750 bp fragments,then ligated into the BamHI site of pBluescript KS(+) and primers T3 and(CT)7 were used for PCR amplification.Positive colonies were screened and plasmids were sequenced.A total of 116 positive colonies were obtained,the sequence testing indicated that 37 of those colonies contained a microsatellite insert and 28 contained full flanking sequence.

Key words: Lasiopodomys brandtii, Small-insert genomic DNA libraries, PCR screen

摘要： 为了研究布氏田鼠(Lasiopodomys brandtii)不同地理种群的遗传差异,克隆其微卫星位点。将布氏田鼠基因组DNA用限制性内切酶消化后的小片段与Pbluescript SK(+)连接,用含有CT重复序列的寡聚核苷酸和质粒上的T3序列作为引物进行PCR扩增,筛选阳性克隆,对质粒上插入位置和片段大小合适的外源DNA序列进行测序。共获得116个阳性克隆,测序结果显示37个包含布氏田鼠微卫星DNA序列,28个具完整的侧翼序列。

关键词: 布氏田鼠, 小片断插入文库, PCR筛选

CLC Number:

S812.6

WANG Deng, SHI Da-zhao. Screening of Small-insert Genomic DNA Libraries for Brandt’s voles by PCR[J]. , 2007, 15(3): 278-282.

王登, 施大钊. PCR方法筛选布氏田鼠小片断插入微卫星文库[J]. , 2007, 15(3): 278-282.

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Screening of Small-insert Genomic DNA Libraries for Brandt’s voles by PCR

PCR方法筛选布氏田鼠小片断插入微卫星文库

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