大蜂螨(Varroa destructor)精氨酸激酶基因的克隆与功能分析

大蜂螨(Varroa destructor)是养蜂业的头号大敌，近年来被认为全球蜂群数量急剧下降的主要原因之一，其遗传背景研究非常薄弱。精氨酸激酶（Arginine Kinase,AK）是节肢动物体内唯一的磷酸原激酶，在能量代谢过程中发挥着重要的作用。本项目拟采用RACE技术和基因步移法分别获得大蜂螨精氨酸激酶基因和基因组序列，应用生物信息学方法对其启动子等调控元件进行分析；同时应用RT-PCR和FISH技术检测AK在大蜂螨不同组织中的分布情况和不同发育时期及不同应急下的基因表达模式；最后从能量代谢系统出发，研究RNA干扰大蜂螨AK基因对其生长发育及繁殖的影响。本项目的开展将从分子水平阐明大蜂螨能量代谢、储存和利用的调节机理，同时为大蜂螨功能基因组研究、蜂螨与蜜蜂协同进化及蜜蜂抗螨基因筛选等研究提供重要数据，为下一步开展RNAi控制蜂螨危害研究奠定基础，进而为全球蜂群的保护作出应有的贡献。

Varroa destructor is the worst enemy of honey bees, it is currently thought to be one of the mainly causes of Collony Collpase Disorder(CCD). So far, the understood of the molecular biology in Varroa destructor was not clear. Arginine kinase (AK) is the only phosphorylase kinase and plays an important role in the energy metabolism in phylum arthropoda. In this project, the AK gene will be cloned from Varroa destructor by RACE, the genomic and protein sequences will be analized with genomic walking and bio-informatics technologies, and the expression of AK gene will be identified in different tissues, different stage and different stimulations by RT-PCR and FISH. In addition, the effects of RNA interference (RNAi) on the energy metabolism in Varroa destructor will be conducted with the double stranded RNA that are designed from the functional region of the AK. As a result, this project will explore the molecular mechanism of energe metabolism, energe storage and energe utilization for Varroa destructor. And it will offer the foundational data to the further researching of functional genes in Varroa destructor. Furthermore, it will contribute to studies of coevolution between varroa mites and their hosted honey bees, mite-resistence genes hunting in honey bee, bio-controlling of varroa destructor based on RNA interference. Thereby, it will contribute to the protection of the global honey bee colonies.