Genetic Analysis of Anopheles mosquitoes

描述（由申请人提供）：昆虫媒介传播的疾病继续给世界上很大一部分人口造成负担。了解负责传播引起这些疾病的病原体的昆虫的基本生物学已经并将继续是制定有效控制战略的必要条件。技术进步常常提供新的机会来扩大我们对基础生物学的理解。该项目将导致为人类疟疾蚊子斯氏按蚊创建增强子和基因陷阱技术，这将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起着至关重要的作用。初步结果表明，转座因子piggyBac在斯氏按蚊基因组中具有高度活性，为构建这些基因技术提供了一个有用的平台。增强子诱捕器和基因诱捕器的原型系统已被创建并引入到斯氏按蚊中。测试两个系统功能的初步筛选已经成功完成。这两种系统在感应增强子和基因方面都是活跃和有效的。描述了一个小的不同的增强阱线集合，它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器和基因诱捕技术将得到改进，以提高效率和有效性。进行增强子和基因陷阱筛选所需的转基因系以及有用的“Gal4驱动系”将成为一种社区资源。斯氏按蚊基因诱捕技术将用于鉴定和分离在各种发育和生理条件下幼虫和成虫唾液腺中特异性表达的基因。该项目将使人们了解唾液腺各种分泌物在蚊虫取食和病原体传播中的重要性，并整合唾液腺结构和功能方面的知识。对人类疾病主要媒介中这一关键组织的新的详细了解，有望导致破坏病原体传播的新策略的构想和实施。

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.