提取CO Ⅰ基因片段鉴定嗜尸性蝇类

Identification of sarcosaphagous flies by amplification and sequence analysis of COⅠ gene fragment from DNA extracts

目的通过扩增蝇类COⅠ基因片段,结合形态学特征鉴定嗜尸性蝇类的种类。方法运用改良平衡酚—tris饱和酚法提取蝇类DNA,进行COⅠ序列扩增和测序,与数据库序列进行比对和分析。结果改良平衡酚—tris饱和酚提取DNA方法可获得有效的蝇类DNA,并可应用于COⅠ基因片段扩增,进而进行蝇类种类的鉴定。结论平衡酚—Tris饱和酚法提取的噬尸性蝇类虫体DNA作为模板,用于COⅠ序列扩增,测序后与数据库目的序列分析比对,可准确鉴定嗜尸性蝇类的种类;和传统的昆虫形态学特征鉴定方法比较,该体系更准确,应用范围更广。

Objective To identify the species of sarcosaphagous flies by amplifying cytochrome oxidase subunitI （COI） gene fragment, combined with morphological characteristics. Methods The DNA of sarcosaphagous flies wasextacted by modified Tris balanced phenol - Tris saturated phenol protocol, the amplificationand sequencing of CO Ifragment were conducted, and the results were then compared with the database for analysis. Results The modifiedDNA extration method by Tris balanced phenol - Tris saturated phenol could obtain effective DNA of sarcosaphagousflies, and be applied for CO I fragment amplification, and thus for identification of the species of sarcosaphagous flies.Conclusion The DNA extracted from sarcosaphagous flies by modified Tris balanced phenol - Tris saturated phenolprotocol could be used astemplate for the amplification of CO I fragment, and the system could identify the speciesof sarcosaphagous flies after sequencing and alignment with the sequences of database. Compared with traditionalidentification methods of using morphological characteristics, the current system is more accurate and could be morewidely applied.