聚合酶链反应（PCR）技术在蚊虫抗性基因研究中的应用

应用ＰＣＲ技术对广泛部分地区的致倦库蚊ＯＰ抗性基因进行测定，实验证明广州地区自然种群和实验室株有相当数量致倦库蚊有扩增的抗性酯酶Ｂ１基因存在。对实验室株及四个地区自然群体的抗性基因进行扩增，发现实验室株阳性率比自然群体为高，四个地区自然群体抗性基因频率分别为２２．５％、２０％、１５％、和７．５％。另外发现雄性库蚊出现的扩增阳性率比雌蚊高。所设计的两对引物均能成功的扩增致倦库蚊抗性基因酯酶Ｂ１，但后

THE STUDY ON THE INSECTICIDE RESISTANCE IN CULEX MOSQUITO BY POLYMERASE CHAIN REACTION(PCR)

Two pairs of primer are designed according to the sequence of esterase B1 gene responsiblefor insecticide organoph0sphorus(OP)resistance in Culex.The OP resistant gene of C.quinquefasciatus in Guangzhou was tested by using Polymerase Chain R eaction(PCR).The result showsthat the positive rate of amplified fragment by primer 2 and 3 is higher than that by primer 1 and2.Insecticide resistant gene of five strains Culex quinquefasciatus from our laboratory,Sipai region,Zhongshan Medical University,Xiaogang region and Zhongshan University were studiedrespectively.The positive rate of laboratory(38%)is the highest,and others are 22.5%,15%,20% and 7.5% respectively.In addition,the higher positive rate in male than in femalewas also observed.In comparison of two different methods of DNA extraction,we feel the simplified method is better because it is not only simple to practise,but also timesaving.Therefore,it might be more suitable for the large scale of investigation in field.PCR and other used methodsof detecting mosquitoes'resistance are also discussed in this paper.