论文部分内容阅读
目的 依据嗜人按蚊和中华按蚊rDNA的ITS2区段基因特征 ,建立一种新的嗜人按蚊和中华按蚊基因鉴别技术。方法 对不同地区的嗜人按蚊、可疑嗜人按蚊和中华按蚊样本通过采用特异性ITS2 引物PCR扩增 ,限制性内切酶RsaI和HinfI消化 ,以及琼脂糖凝胶电泳分析进行PCR -RFLP基因鉴别。 结果 不同地区嗜人按蚊rDNA的ITS2 基因PCR扩增产物能被限制性内切酶HinfI酶切 ,并显示一条 4 5 0bp的酶切DNA条带 ;中华按蚊的ITS2 基因PCR扩增产物则能被限制性内切酶RsaI酶切 ,并显示 4 0 0bp和 2 0 0bp两条酶切DNA条带 ;辽宁可疑嗜人按蚊的PCR -RFLP结果与嗜人按蚊相同而广东珠海可疑嗜人按蚊的PCR -RFLP结果与中华按蚊相同。结论 依据rDNA的ITS2 区段基因特征建立的PCR -RFLP技术可用于嗜人按蚊和中华按蚊的基因鉴别。采用该PCR -RFLP基因鉴别技术发现辽宁可疑嗜人按蚊的基因与中国大陆的嗜人按蚊属同种 ,而广东可疑嗜人按蚊的基因与中华按蚊属同种。
Objective To establish a new Anopheles anthropophagus and Anopheles sinensis genetic identification technique based on ITS2 gene of Anopheles anthropophagus and Anopheles sinensis. Methods The An. Anthropophagus, Susceptible An. Anopheles and An. Sinensis samples were digested by specific ITS2 primers, restriction endonucleases RsaI and HinfI, and analyzed by agarose gel electrophoresis. RFLP gene identification. Results PCR amplification products of ITS2 gene of Anopheles anthropophagus in different areas were digested by restriction endonuclease HinfI and showed a 455bp digested DNA band. ITS2 gene of Anopheles sinensis was amplified by PCR Can be digested with restriction endonuclease RsaI, and showed 400bp and 200bp DNA digestion bands; PCR-RFLP results of Liaoning suspicious Anopheles anthropophagus is the same as Anopheles anthropophagus and suspicious addicts of Zhuhai, Guangdong Anopheles mosquito PCR -RFLP results with the same Anopheles sinensis. Conclusion The PCR-RFLP technique based on the ITS2 gene of rDNA can be used to identify the Anopheles anthropophagus and Anopheles sinensis. Using this PCR-RFLP gene identification technique, it was found that the genes of Anopheles anthropophagus and Liaoning Anopheles mosquitoes were the same species in mainland China, while the genes of Anopheles anthropophagus in Guangdong were the same species as Anopheles sinensis.