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应用PCR-RFLP技术分析斑翅果蝇及其相关种间的差异。选用果蝇COI基因通用引物并通过PCR扩增斑翅果蝇、亚艳丽果蝇等11种15个样品COI基因序列,对PCR产物进行回收和测序,根据测序结果及NCBI上序列比对结果,选用限制性内切酶ApaⅠ和HinfⅠ分别对果蝇COI基因PCR产物进行酶切,酶切产物经琼脂糖电泳分析。结果表明:同时使用ApaⅠ和HinfⅠ酶,可以将斑翅果蝇与其他种相区分。根据果蝇COI基因建立的PCR-RFLP技术可对斑翅果蝇快速、准确进行鉴定。
The PCR-RFLP technique was used to analyze the differences between Drosophila melanogaster and its related species. The common COI gene primers of Drosophila melanogaster were selected and the COI gene sequences of 11 kinds of samples including 11 species of Drosophila melanogaster and Asian fruit flies were amplified by PCR. The PCR products were recovered and sequenced. According to the sequencing results and sequence comparison results of NCBI, Restriction endonucleases Apa Ⅰ and Hinf Ⅰ were used to digest the Drosophila COI gene PCR products respectively. The digested products were analyzed by agarose gel electrophoresis. The results showed that Drosophila melanogaster could be distinguished from other species by using both ApaI and HinfI enzymes. According to the PCR-RFLP technology established by Drosophila COI gene, Drosophila melanogaster can be rapidly and accurately identified.