Glutathione-s-transferase mediated pyrethroid resistance in Culex pipiens s.l.

说明（由申请人提供）：拟除虫菊酯类杀虫剂是美国和非洲用于控制库蚊种群的主要杀虫剂。这些蚊子种群正在出现对拟除虫菊酯的抗药性，它们在美国是西尼罗病毒的载体，在非洲是裂谷热病毒和班克罗夫特丝虫病的载体。直接证据强烈表明，这种抗性是由酶解毒介导的，包括谷胱甘肽S转移酶（GSTs）。因此，本研究的目的是分离负责介导抗性的GST同工酶，以便开发诊断抗性状态的检测方法。为实现既定目标，将追求以下目标：目标# 1。产生单独的对拟除虫菊酯耐药的成蚊和幼蚊。纯化并获得将谷胱甘肽与拟除虫菊酯结合的gst的氨基酸序列。使用细菌表达系统Aim #4克隆和表达GST同工酶。开发基于GST的拟除虫菊酯耐药性诊断分析方法，一旦这些GST同工酶被确定并开发出分析方法，它们就可用于耐药性监测、缓解和管理战略，以延长拟除虫菊酯的有效使用。这些研究还将有助于了解代谢抗性的机制，以帮助开发可能增强拟除虫菊酯类杀虫剂的新型、下一代杀虫剂和增效剂。

DESCRIPTION (provided by applicant): Pyrethroids are the major insecticides used to control populations of Culex pipiens s.l. mosquitoes in the USA and Africa. Resistance to pyrethroids is emerging in populations of these mosquitoes, where they are vectors of West Nile virus in the USA and Rift Valley fever virus and Bancroftian filariasis in Africa. Direct evidence strongly suggests that this resistance is mediated by enzymatic detoxification including Glutathione S Transferases (GSTs). The goal of this research is therefore to isolate those GST isozymes responsible for mediating resistance for the purpose of developing assays for diagnosing the status of resistance. To achieve the stated goal the following aims will be pursued; Aim #1. Generate separate adult and larval pyrethroid resistant strains of Culex pipiens complex mosquitoes Aim #2. Purify and obtain amino acid sequence of GSTs that conjugate glutathione to pyrethroids Aim #3. Clone and express GST isozymes using a bacterial expression system Aim #4. Develop GST based diagnostic assays for pyrethroid resistance Once these GST isozymes are identified and assays developed they can be used in resistance monitoring and mitigation and management strategies to prolong the effective use of pyrethroids. These studies will also lead to understanding the mechanism of metabolic resistance to aid in development of novel, next generation insecticides and synergists that may augment pyrethroids.