Molecular and functional characterization of olfaction genes in tsetse flies

采采蝇嗅觉基因的分子和功能特征

基本信息

批准号：
8411357
负责人：
Serap AKSOY
金额：
$ 5.67万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-03-20 至 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8411357
关键词：
Address Adult Africa Africa South of the Sahara African African Trypanosomiasis Agriculture Animals Applied Genetics Area Behavior Behavioral Bioinformatics Biology Collaborations Communicable Diseases Communities Companions Cues DNA Data Data Analyses Databases Development Diagnostic Disease Disease Vectors Drug resistance Ecology Educational workshop Environmental Wind Exhibits Gene Expression Profile Gene Family Gene Silencing Genes Genetic Genetic Vectors Genome Genomics Goals Government Agencies Grant Head Health Heterogeneity Human Human Activities In Vitro Information Systems Insecta Institution International Kenya Lead Life Literature Mediating Medical Mentors Methodology Methods Molecular Molecular Profiling Nature Odors Parasites Pharmaceutical Preparations Physiological Physiology Population Population Biology Process Proteins Public Health Publishing RNA RNA Interference Reproduction Research Research Activity Research Institute Research Personnel Resources Risk Role Sahara Sampling Scientist Site Smell Perception Sudan Tanzania Techniques Technology Testing Tissues Training Transcript Tropical Disease Trypanosoma Trypanosoma brucei rhodesiense Trypanosomiasis Tsetse Flies Uganda Vaccines Work World Health Organization base comparative disorder control epidemiological model fly genome sequencing human disease improved interdisciplinary approach international center new technology next generation sequencing odorant-binding protein parent grant programs research and development response tool transmission process vector vector control vector transmission

项目摘要

DESCRIPTION (provided by applicant): The overall goal of this study is to provide information on molecular processes in tsetse vectors that mediate their olfactory processes, enabling them to locate their hosts and other critical resources for their survival and reproduction. The goals will be achieved by prospecting for and validating olfaction responsive genes in selected tsetse species (Glossina pallidipes, Glossina austeni and Glossina fuscipes fuscipes) of medical and veterinary importance in Kenya. We propose an integrated and interdisciplinary approach to: 1) Determine differences in olfactory responsive genes in sympatric Glossina pallidipes and Glossina austeni in Shimba hills and Glossina fuscipes fuscipes in Mbita in Kenya and 2) Determine functional roles of olfactory responsive genes in sympatric G. pallidipes and G. austeni in Shimba hills and G. f. fuscipes in Mbita districts in Kenya. We hypothesize that putative olfactory genes among these flies are orthologuous, differentially expressed and significantly influence responses of the flies to different odors. Adults of the selected tsetse species will be sampled from respective sites and RNA will be extracted from head tissues. We will undertake a discovery project by using next generation sequencing technologies to determine global transcriptome-level heterogeneities exhibited between the different species. The DNA obtained from the gut contents of flies will be subjected to PCR amplification with COI marker to determine the putative host species from Barcode of Life Data systems (BOLD) database. Following the comparative transcriptome analysis from the different species we will in vitro validate the presence and expression profiles of select set of putative olfaction related genes through qPCR. The functional role(s) of selected putative olfaction genes will be further investigated through RNAi gene silencing methodologies in conjunction with electro-antennogram and Wind-Tunnel techniques to establish the roles of these genes in modulating the physiology and behavior of the flies. Availability of comprehensive information on olfaction related genes, and their respective functional roles in modulating tsetse olfactory behavior will facilitate further R&D to exploit these genes to control tsetse fly vectors. This research will be done primarily in Kenya at International Centre of Insect Physiology Ecology and Kenya Agricultural Research Institute - Trypanosomiasis Research Center in collaboration with Paul Mireji with companion grant being Global Infectious Diseases (GID) D43 TW007391 (08/01/2008- 03/31/2013).

描述（由申请人提供）：这项研究的总体目标是提供有关采用辅助过程的采采媒体中分子过程的信息，从而使他们能够找到其宿主和其他关键资源以生存和繁殖。肯尼亚的医学和兽医重要性，将通过探明和验证选定采集物种（Glossina Pallidipes，Glossina Austeni和Glossina Fuscipes fuscipes）中的嗅觉反应基因来实现目标。我们提出了一种综合和跨学科的方法：1）确定在Shimba Hills中的胶质胶质体和Glossina Austeni中的嗅觉响应基因的差异和肯尼亚MBITA中的格纹液的差异，并确定肯尼亚和2）在同对基因的功能和2）确定Sympatric gel.pallip的功能。 Hills和G. f。肯尼亚Mbita地区的fuscipes。我们假设这些苍蝇中假定的嗅觉基因是直系同源的，差异表达的，并且显着影响苍蝇对不同气味的反应。将从各个部位取样所选采采种物种的成年人，将从头组织中提取RNA。我们将通过使用下一代测序技术来确定不同物种之间表现出的全球转录组异质性。从苍蝇的肠道含量获得的DNA将通过COI标记进行PCR扩增，以确定Life Data Systems（BOLD）数据库条形码的假定宿主物种。根据来自不同物种的比较转录组分析，我们将在体外验证通过qPCR选择一组推定嗅觉相关基因的存在和表达谱。将通过RNAi基因沉默方法与电抗验证函数和风态技术技术一起进一步研究选定推定嗅觉基因的功能作用，以确定这些基因在调节果蝇的生理学和行为中的作用。有关相关基因的综合信息及其在调节采采嗅觉行为中的各自功能作用将有助于进一步的研发，以利用这些基因来控制TSETSE Fly Vectors。这项研究将主要在肯尼亚国际昆虫中心进行生理生态学和肯尼亚农业研究所 - 锥虫病研究中心与Paul Mireji与Paul Mireji合作，同伴Grant是全球传染病（GID）D43 TW007391（08/01/2008- 2008- 03/03/31/2013）。