目的 确定布氏田鼠细胞色素C氧化酶亚基Ⅰ (CO Ⅰ)基因巢式PCR扩增的方法.方法 采集内蒙古锡林郭勒盟阿巴嘎旗青格勒宝拉格地区的布氏田鼠12只,取肝脏组织,提取DNA,采用巢式PCR方法扩增CO Ⅰ基因,并对扩增产物进行测序.结果 巣氏PCR内外引物分别扩增出657、1 132bp的产物,与目的片段大小一致.内引物PCR扩增共获得12条布氏田鼠CO Ⅰ基因序列片段,测序均为CO Ⅰ基因片段,在GenBank中序列号为KF182196-KF182207;12条CO Ⅰ基因序列进行比对,相似性为100%,无碱基的变异.结论 巢氏PCR方法可有效扩增出布氏田鼠COⅠ基因,避免产生假基因现象.
Objective To determine a method for amplification of cytochrome C oxidase subunit Ⅰ (CO Ⅰ) gene of Brandt's vole.Methods 发he Brandt's Voles were captured in Abagaqi Xilingol League Inner Mongolia,and DNA was extracted from liver tissue.CO Ⅰ gene was amplified by nested PCR and sequenced afterwards.Results A band of 657 bp and 1 132 bp was amplified by internal and external PCR primers,respectively,which were consistent with expected sizes.A total of 12 segments of Brandt's Vole CO Ⅰ gene sequences were amplified by PCR and verified by sequencing.一he sequence number was KF182196-KF182207 in GenBank.After gene sequence alignment of the 12 CO Ⅰ gene sequences,it was found that the similarity was 100%,and no base mutation.Conclusion CO Ⅰ gene of Brandt's Vole could be amplified by nested PCR without pseudo gene.