Author
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Wei Pridgeon, Yuping |
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Becnel, James |
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Clark, Gary |
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Linthicum, Kenneth - Ken |
Submitted to: Journal of Medical Entomology
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 3/11/2009 Publication Date: 5/1/2009 Citation: Pridgeon, Y.W., Becnel, J.J., Clark, G.G., Linthicum, K. 2009. Permethrin induces overexpression of multiple genes in Aedes aegypti. Journal of Medical Entomology. 46(3):580-587. Interpretive Summary: In order to understand the response of mosquitoes to pesticide treatment at molecular level, subtractive cDNA hybridization technique was used to isolate genes that are overexpressed in response to permethrin treatment. Eighteen different genes were isolated from a permethrin-treated vs acetone-treated Aedes aegypti subtractive library. Real-Time PCR results showed that eight of the 18 gene’s transcriptional levels in permethrin-treated Ae. aegypti were at least 2- fold higher than that in acetone-treated Ae. aegypti. These eight genes include three functionally known genes (cytochrome c oxidase subunit III, NADH2 dehydrogenase, deltamethrin resistance associated protein), three functionally unknown genes (Ae. aegypti putative 16.9 kDa secreted protein, Anopheles gambiae ENSANGP00000019508, Cryptococcus neoformans hypothetical protein CNE05340) and two novel genes. Transcriptional levels for 11 of the 18 genes were induced significantly higher by permethrin than by fipronil. Overexpression of multiple genes induced by permethrin suggests that they might play functional roles in response to pesticide treatment and/or initial steps of insecticide resistance development. Technical Abstract: Using the PCR-select subtractive cDNA hybridization technique, 18 different genes were isolated from a permethrin-treated vs acetone-treated Aedes aegypti subtractive library. QPCR results revealed that eight of the 18 gene’s transcriptional levels in permethrin-treated Ae. aegypti were at least 2- fold higher (ranging from 2.6'0.5 to 4.8'0.2) than that in acetone-treated Ae. aegypti. These eight genes include three functionally known genes (cytochrome c oxidase subunit III, NADH2 dehydrogenase, deltamethrin resistance associated protein), three functionally unknown genes (Ae. aegypti putative 16.9 kDa secreted protein, Anopheles gambiae ENSANGP00000019508, Cryptococcus neoformans hypothetical protein CNE05340) and two novel genes. Transcriptional levels for 11 of the 18 genes were induced significantly higher by permethrin than by fipronil (p<0.05). Our results suggest that subtractive cDNA hybridization and QPCR are powerful techniques to identify differentially expressed genes in response to pesticide treatment. Overexpression of multiple genes induced by permethrin suggests that they might play functional roles in response to pesticide treatment and/or initial steps of insecticide resistance development. |