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大仓鼠DNA 指纹谱探针的筛选

宋铭晶  徐来祥  张知彬   

  1. 中国科学院动物研究所农业虫鼠害综合治理国家重点实验室, 北京
  • 出版日期:2005-10-30 发布日期:2008-07-07

Screening of the DNA Fingerprint Probes of Greater Long-tailed Hamster( Tscherskia triton)

SONG Mingjing,XU Laixiang,ZHANG Zhibin   

  • Online:2005-10-30 Published:2008-07-07

摘要: 采用一种简便提取高质量DNA 的方法, 从大仓鼠肝脏组织中提取其总DNA , 分别以人工合成的微卫星核心序列(GTG)5和(CA)8做单一引物, 进行特异引物PCR 反应。电泳检测后回收15 条特异性片段。与被标记过的大仓鼠基因组DNA 反向杂交结果表明, 15 个片段中(GTG)5-8 、(CA)8-1b和(CA) 8-5b产生了较强的阳性信号。我们依据3 个片段的测序结果设计适合DIG标记的探针, 该探针得到的大仓鼠不同地理种群个体的指纹图谱有较高的个体特异性和种群多态性, 而且与传统的来源于其它生物重复序列的探针如33.6 和33.15 形成的指纹图谱相比得到的变异适中, 便于统计。

关键词: 大仓鼠, DNA 指纹谱, 探针, 微卫星, 反向杂交

Abstract: We extracted the genomic DNA from liver tissues of Tscherskia triton using isolation of high quality DNA method. Then the DNA template of Tscherskia triton was amplified with a single primer composing ( GTG)5or (CA)8 by using the polymerase
chain reaction (PCR). After agarose gel electrophoresis, fifteen specific amplification fragments were retrieved. Among these specific PCR products, we found that fragment (GTG)5 - 8,(CA)8 - 1b and (CA)8 - 5b showed more positive signals than others in reverse hybridization test with genomic DNA. We designed the DIG labeled probes according to the sequencing result of these three fragments. The DNA fingerprints pattern of the greater long-tailed hamster individuals from different geographic areas hybridized by these probes were individual-specific and population-polymorphic. Moreover in contrast to DNA fingerprints hybridized by conventional probes derived from other sources, such as 33.6 and 33.15, these probes detected medium level of variation in DNA fingerprints, which were easier to score.

Key words: DNA fingerprint, Greater Long2tailed hamster ( Tscherskia triton), Microsatellite, Probe, Reverse hybridization