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United States Department of Agriculture

Agricultural Research Service

Research Project: CONTROL AND PROTECTION TOOLS FOR INTEGRATED PEST MANAGEMENT OF MOSQUITOES AND FILTH FLIES

Location: Mosquito and Fly Research Unit

Title: Identification of genes differentially expressed during heat shock treatment in Aedes aegypti.

Authors
item Zhao, Liming
item Wei Pridgeon, Yuping
item Becnel, James
item Clark, Gary
item Linthicum, Kenneth

Submitted to: Journal of Medical Entomology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: January 2, 2009
Publication Date: April 30, 2009
Citation: Zhao, L., Pridgeon, J. W., Becnel, J. J., Clark, G. G., Linthicum, K. J. 2009.Identification of genes differentially expressed during heat shock treatment of Aedes aegypti. Journal of Medical Entomology. 46(3):490-495.

Interpretive Summary: To understand high temperature alteration in gene expression of a vector mosquito and develop a novel control approach, we use suppression subtractive hybridization (SSH) to identify target transcripts to heat shock treatment in female Aedes aegypti. Heat shock treatment of adult Ae. aegypti was carried out for 1 hr at 42 ºC. According to our data, several genes of heat shock protein (HSP) families are known to be expressed in mosquitoes and may be crucial in responding to stress induced by elevated temperature. To determine other genes expressed in response to heat stress, we used the powerful technology---PCR-based cDNA subtraction. Target transcripts up-regulated by heat shock included 5 different HSP gene families as well as 27 other genes, such as serine-type endopeptidase, cytochrome c oxidase and glutamyl aminopeptidase. Additionally, some novel genes, cytoskeleton and ribosomal genes were found to be differentially expressed, and three novel up-regulated sequences belonging to a low-abundance class of transcripts were obtained. Up-regulated/down-regulated transcripts from heat shock treatment were further confirmed and quantified by quantitative real-time PCR. . Identification of these groups of gene families may provide critical information needed for designing novel control strategies for medically important disease vectors and identifying new pathways to target for the development of genetic molecular pesticides.

Technical Abstract: : Temperature is important for mosquito development and physiological response. Several genes of heat shock protein (HSP) families are known to be expressed in mosquitoes and may be crucial in responding to stress induced by elevated temperature. Suppression subtractive hybridization (SSH) was used to identify target transcripts to heat shock treatment in female Aedes aegypti. SSH is a method in which two populations of mRNA are compared to obtain clones of genes expressed in one population but not in the other. Subtraction was performed in both directions enriching for cDNAs differentially expressed between non heat shock control and heat shock treatment. Heat shock treatment of female Ae. aegypti was carried out for 1 hr at 42 ºC. Clones from differentially expressed genes were evaluated by sequencing. Target transcripts up-regulated by heat shock included 5 different HSP gene families as well as 27 other genes, such as cytochrome c oxidase, serine-type endopeptidase, and glutamyl aminopeptidase. Additionally, some novel genes, cytoskeleton and ribosomal genes were found to be differentially expressed, and three novel up-regulated sequences belonging to a low-abundance class of transcripts were obtained. Up-regulated/down-regulated transcripts from heat shock treatment were further confirmed and quantified by quantitative real-time PCR. High temperatures can alter the gene expression of a vector mosquito population, and further characterization of these differentially expressed genes will provide information useful in understanding the genetic response to heat shock treatment, which can then be used to develop novel approaches to genetic control.

Last Modified: 8/31/2014
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