摘要
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。
Aim To establish a new genetic identification technique based on the genetic characteristics of ribosomal DNA ITS2 region of Anopheles anthropophagus and Anopheles sinensis Methods The PCR-RFLP technique has been used for genetic identification of Anopheles anthropophagus and Anopheles sinensis The ribosomal DNA ITS2 regions of Anopheles anthropophagus from different areas,suspected Anopheles anthropophagus and Anopheles sinensis have been amplified then digested with Rsa I and Hinf I and analyzed on an agarose gel Results The ribosomal DNA ITS2 region PCR products of Anopheles anthropophagus from different areas could be digested with Hinf I and showed a 450bp DNA bane and the same result has been obtained from suspected Anopheles anthropophagus from Liaoning province while PCR products from Anopheles sinensis ITS2 region could be digested with Rsa I and showed 400bp and 200bp banes and the suspected Anopheles anthropophagus from Guangdong province showed the same result Conclusion The established PCR RFLP technique based on the genetic characteristics of ribosomal DNA ITS2 region can be used for genetic identification of Anopheles anthropophagus and Anopheles sinensis The genetic analysis showed that the suspected Anopheles anthropophagus from Liaoning province is same as Anopheles anthropophagus while the suspected Anopheles anthropophagus from Guangdong province is same as Anopheles sinensis
出处
《中国人兽共患病杂志》
CSCD
北大核心
2002年第6期39-42,共4页
Chinese Journal of Zoonoses
