Expression analysis and knockdown of two antennal odorantbinding protein genes in Aedes aegypti.pdfVIP

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Expression analysis and knockdown of two antennal odorantbinding protein genes in Aedes aegypti.pdf

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Expression analysis and knockdown of two antennal odorantbinding protein genes in Aedes aegypti

Journal of Insect Science: Vol. 10 | Article 171 Sengul and Tu Journal of Insect Science | 1 Expression analysis and knockdown of two antennal odorant- binding protein genes in Aedes aegypti Meryem S. Sengul1,2a and Zhijian Tu1b 1Department of Biochemistry, Virginia Polytechnic Institute and State University, Blacksburg, VA, 24061, USA 2Current address: Department of Biology, Bozok University, Yozgat, 66200, Turkey Abstract The presence and expression of odorant-binding proteins (OBPs) in the olfactory organs suggest that they play an important role in mosquito olfaction. However, no direct evidence has been found for their involvement in the host-seeking behavior of mosquitoes. It is important to establish a method in which a loss-of-function test can be performed to determine the possible role of these genes in olfaction. In this study, a double subgenomic Sindbis virus expression system was used to reduce the expression of two Obp genes in Aedes aegypti L (Diptera: Culicidae), AaegObp1 and AaegObp2. Quantitative real-time PCR analysis showed predominant expression of both genes in the female antennae, the primary olfactory tissue of mosquitoes. Moreover, at 11 days post virus-inoculation, the mRNA levels of AaegObp1 and AaegObp2 were significantly reduced in olfactory tissues of recombinant virus-inoculated female mosquitoes compared to that of controls by approximately 8 and 100-fold, respectively. These data suggest that the double subgenomic Sindbis virus expression system can be efficiently used to knockdown Obp gene expression in olfactory tissues of mosquitoes. We discuss the potential for a systematic analysis of the molecular players involved in mosquito olfaction using this newly developed technique. Such analysis will provide an important step to interfere with the host- seeking behavior of mosquitoes to prevent the transmission of diseases. Keywords: Gene knockdown, mosquito, olfaction, RNAi, Sindbis virus Abbreviations: AaegObp1, Aedes