Genetic Analysis of Anopheles mosquitoes

描述(申请人提供)：昆虫传播的疾病继续给世界上很大一部分人口带来负担。了解传播引起这些疾病的病原体的昆虫的基本生物学一直是并将继续是制定有效的控制战略的关键。技术进步经常提供新的机会来扩大我们对基础生物学的理解。该项目将为人类疟疾蚊子斯氏按蚊创造增强子和基因陷阱技术，使研究人员能够以新的和强大的方式识别、分离和分析基因。这项技术将应用于唾液腺的研究，唾液腺在疟原虫等病原体的传播中发挥着至关重要的作用。初步结果表明，转座元件iggyBac在斯氏按蚊基因组中高度活跃，使其成为建立这些基因技术的有用平台。已经建立了原型增强子陷阱和基因陷阱系统，并将其引入斯氏按蚊。测试这两个系统功能的初步筛选工作已经顺利完成。这两个系统在分别检测增强剂和基因方面都是活跃和有效的。描述了一个小而多样的增强子-陷阱线集合，这提供了一个原理证明。在原型设计的基础上，现有的斯氏按蚊增强子和基因捕获技术将进行改进，以提高效率和效果。进行增强子和基因陷阱筛选所需的转基因品系，以及有用的“Gal4驱动品系”将成为社区资源。斯氏按蚊基因捕获技术将被用来鉴定和分离在各种发育和生理条件下在幼虫和成虫唾液腺中特异表达的基因。该项目将使人们能够了解各种唾液腺分泌物在蚊子喂养和病原体传播中的重要性，并整合有关唾液腺结构和功能的知识。这种对人类疾病主要媒介中这一关键组织的新的详细了解，有望导致中断病原体传播的新战略的构思和实施。

DESCRIPTION (provided by applicant): Insect vector-borne diseases continue to burden a large fraction of the world's population. Understanding the basic biology of the insects responsible for transmitting the pathogens that cause these diseases has been and will continue to be essential for the development of effective strategies of control. Technological advances often provide novel opportunities to enlarge our understanding of basic biology. This project will result in the creation of enhancer- and gene-trap technologies for the human malaria mosquito Anopheles stephensi that will enable researchers to identify, isolate and analyze genes in new and powerful ways. The technology will be applied to the study of the salivary glands, an organ that plays a vital role in the transmission of pathogens such as Plasmodium. Preliminary results show that the transposable element piggyBac is highly active within the genome of Anopheles stephensi, making it a useful platform upon which to build these genetic technologies. Prototype enhancer- trap and gene-trap systems have been created and introduced into Anopheles stephensi. Preliminary screens to test the functionality of both systems were successfully completed. Both systems are active and effective at sensing enhancers and genes, respectively. A small diverse collection of enhancer-trap lines is described which provides a proof of principle. Based on prototype designs, existing Anopheles stephensi enhancer- and gene-trap technologies will be modified for increased efficiency and effectiveness. The transgenic lines necessary for conducting enhancer- and gene-trap screens, as well as, useful 'Gal4 driver lines' will become a community resources. Anopheles stephensi gene-trap technology will be used to identify and isolate genes specifically expressed in the salivary glands of larvae and adults under a variety of developmental and physiological conditions. This project will enable the importance of various salivary gland secretions in mosquito feeding and pathogen transmission to be understood, and to integrate knowledge concerning salivary gland structure and function. This new detailed understanding of this critical tissue in a major vector o human disease is expected to lead to the conception and implementation of new strategies for disrupting pathogen transmission.