大仓鼠DNA指纹谱探针的筛选

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

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。

We extracted the genomic DNA from liver tissues of Tscherskia triton usiog isolation of high quality DNA method. Then the DNA template of Tscherskia triton was amplified with a single primer composing （GTG）5 or （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 rat-like 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.