虻虫的PCR-RFLP鉴别研究

Molecular Identification of Medicinal Gadfly and Its Adulterants by PCR-RFLP Analysis

目的:建立简单、准确的虻虫DNA分子标记鉴别方法。方法:扩增并分析虻虫及7种常见伪品的细胞色素C氧化酶I亚基基因(CO I)序列,根据差异片段设计聚合酶链反应-限制性内切酶酶切长度多态性(PCR-RFLP)引物,使用限制性内切酶进行酶切鉴别。结果:引物对Meng Chong-Dig.F/Meng Chong-Dig.R可扩增虻虫及其伪品CO I序列,产生约490 bp条带,限制性内切酶Dra I可以识别虻虫及其伪品的差异序列,仅虻虫的CO I片段可被酶切形成两个片段,从而特异性鉴别是否为虻虫。结论:本实验建立的PCR-RFLP可以作为虻虫的DNA鉴定方法。

Objective： To establish a convenient and effective DNA molecular identification method for medicinal gadfly. Methods ： CO I sequences of gadfly and 7 adulterants were amplified and analyzed, PCR-RFLP primers were designed based on differential fragment, and restriction enzyme digest was further conducted to identify gadfly and the 7 adulterants. Results： Approximately 490 bp specific fragment bands from medicinal gadfly and its adulterants were amplified by MengChong-Dig. F/MengChong-Dig. R. Dra I restriction endonuclease could only digest the amplified fragment of medicinal gadfly into two smaller pieces, therefore identifying medicinal gadfly specifically. Conclusion： The methods of PCR-RFLP developed in this research is an accurate identification method for medicinal gadfly.