Hermes Element Transposition in Mosquitoes

蚊子中的赫耳墨斯元素转位

• 批准号: 7010012
• 负责人: PETER William ATKINSON
• 金额: $ 41.79万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7010012
• 关键词: Aedes; animal population genetics; arthropod genetics; dimer; disease vectors; genetically modified animals; laboratory mouse; site directed mutagenesis; transfection /expression vector; transposon /insertion element

DESCRIPTION (provided by the applicant): The long-term goal of this project is to understand the behavior of introduced transposable elements in transgenic mosquitoes that are vectors of human disease. It is the central proposition of the research described herein that the ability of a transposable element to be used both as an efficient genetic tool in the laboratory in mosquitoes, as well as an effective "gene driving mechanism" of beneficial effector genes in the field is determined by both how the transposable element functions, and how it interacts with the new host mosquito genome. This proposition will be examined using the Hermes transposable element, which is a member of the hAT family of transposable elements, and which is an effective transformation vector of the mosquito, Aedes aegypti. The Specific Aims of this proposal are thus: 1/ to purify and characterize the Hermes transposase, 2) to understand the structure: function relationships of the Hermes element, 3) to examine interactions of the Hermes transposase, and 4) to isolate hyperactive mutants of Hermes transposase and to test them for hyperactivity in mosquitoes. Purified biologically active Hermes transposase will be used to determine the optimal physical and biological parameters for Hermes element excision and transposition and for the role that host factors may play in Hermes element transposition in mosquitoes. Regions of the both the transposase and the element, such as the inverted terminal repeats and adjacent conserved motifs, that are required for transposition will be identified and, using mutagenesis, their precise roles in this process determined. The role of circular forms of the Hermes element, which we have identified to be present in transgenic insects containing autonomous forms of Hermes element will be investigated. Finally, both eukaryotic and prokaryotic based expression systems, both of which produce active Hermes transposase, will be used to identify hyperactive forms of the Hermes element system that will then be tested for elevated levels of activity in Ae. aegypti. To date we know little about how the transposable elements used to genetically transform mosquitoes either work or behave in mosquito genomes. This now constitutes the most significant bottleneck to the extension of transposable element-based technologies into medically important mosquito species. This technology will complement existing approaches to the control of mosquito-borne disease control and will provide new avenues with which to approach urgent problems arising from the increase in the geographical distribution of these devastating diseases which impact both human health and national economies. This new technology will enable mosquito genes required for pathogen transmission to be identified and isolated, will improve the efficiency with which transgenic mosquito strains can be identified, and will also provide important data concerning the likelihood that genes that prevent pathogen transmission can be spread through mosquito populations by transposable elements.

描述（由申请人提供）：该项目的长期目标是了解转基因蚊子中引入的转座元素的行为，这些蚊子是人类疾病的媒介。本文描述的研究的核心主张是，在蚊子实验室中，可以用作有效的遗传工具的能力，以及该领域中有益效应基因的有效“基因驱动机制”，由可转移元素功能以及如何与新宿主蚊子基因组相互作用来确定。该命题将使用爱马仕（Hermes）转座元素进行检查，后者是可转移元素的帽子家族的成员，它是蚊子埃德斯伊斯特（Aedes aegypti）的有效转化向量。因此，该提案的具体目的是：1/要净化和表征爱马仕转座酶，2）理解结构：爱马仕元素的功能关系，3）检查爱马仕转座酶的相互作用，以及4）4）以分离爱马仕转培酶的过度活跃突变体，并测试它们在蚊子中的过度活性。纯化的生物活性爱马仕转座酶将用于确定HERMES元素切除和转座的最佳物理和生物学参数，以及宿主因子在HERMES元素中可能在蚊子中发挥作用的作用。将识别转座酶和元素的区域，例如倒末端重复序列和相邻的保守基序，将被鉴定为转置所需的区域，并使用诱变，在确定的过程中确定其精确的作用。将研究爱马仕元素的圆形形式的作用，我们已经确定在包含爱马仕元素的自主形式的转基因昆虫中存在。最后，基于真核和原核的表达系统，两者都会产生活跃的爱马仕转座酶，用于鉴定爱马仕元素系统的多动激活形式，然后将测试AE中AE的活性水平升高。埃及。 迄今为止，我们对用于转化蚊子在蚊子基因组中起作用或作用的可转移元素如何了解。现在，这构成了将基于元素的技术扩展到医学重要的蚊子物种中最重要的瓶颈。这项技术将补充现有的方法来控制蚊子 - 传播疾病的控制，并将提供新的途径，以解决这些毁灭性疾病的地理分布增加，这会影响人类健康和国民经济。这项新技术将使病原体传播所需的蚊子基因被鉴定和隔离，将提高可以鉴定的转基因蚊子菌株的效率，并且还将提供有关防止病原体传播的基因可以通过可通过转换元素通过蚊子种群传播的重要数据。