巢式PCR扩增布氏田鼠细胞色素C氧化酶亚基Ⅰ基因

Amplification of cytochrome C oxidase subunit Ⅰ gene of Brandt＆#39;s Vole by nested PCR

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 The Brandt＇s Voles were captured in Ahagaqi 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. The 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 he amplified by nested PCR without pseudo gene.