Expression analysis and knockdown of two antennal odorant-binding protein genes in Aedes aegypti.
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.
Department of Biochemistry, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, USA. senay.sengul@bozok.edu.tr
Meryem S
MS
Sengul
Zhijian
Z
Tu
D000330:Aedes / Q000235:genetics* / Q000378:metabolism; D000704:Analysis of Variance; D000818:Animals; D057506:Arthropod Antennae / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors / Q000235:genetics; D007300:Insect Control / Q000379:methods*; D018035:Receptors, Odorant / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012845:Sindbis Virus
D017931:DNA Primers; D018035:Receptors, Odorant; C045648:odorant-binding protein