殺虫剤抵抗性解析用DNAマイクロアレイの構築と検証

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A microarray containing PCR products from a large number of candidate genes in insecticide resistance was constructed. A total of 646 genes putatively involved in insecticide resistance and positive control genes were obtained from six insect species, Apis cerana, Blattella germanica, Spodoptera exigua, Musca domestica, Nila-parrata lugens and Culex pipiens. These genes represent cytochrome P450, ion channel, protective enzyme and other genes, which may play important roles in insecticide resistance. The optical microarray printing concentration (250ng/μl) was determined using gradient concentration hybridization assay. The DNA microarray was then constructed with all target genes. A PCR probe composed of 18 target genes was prepared to validate the hybridization specificity of the microarray. The positive rate of this validation test was 83.3%. In addition, analysis of differential gene expression between susceptible and multi-resistant (cypermethrin and malathion) strains of M. domestica showed that 17 genes were over-expressed in the resistant strain. The results imply that cytochrome P450 genes, especially the genes of CYP4 and 6 families, play important roles in the resistance of M. domestica to both insecticides. The 17 genes validated by using qRT-PCR were found to have a similar tendency, mostly compared with using a microarray, indicating that this microarray is suitable for studying insecticide resistance.
日本農薬学会の論文
2007-02-20

殺虫剤抵抗性解析用DNAマイクロアレイの構築と検証

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