Hermes Element Transposition in Mosquitoes
蚊子中的赫耳墨斯元素转位
基本信息
- 批准号:6844705
- 负责人:
- 金额:$ 42.87万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 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:
- 关键词:
项目摘要
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作为人类疾病媒介的转基因蚊子中引入的转座子的行为。这里描述的研究的中心命题是,转座元件在实验室用作蚊子的有效遗传工具的能力,以及在田间作为有益效应基因的有效“基因驱动机制”的能力,既取决于转座元件的功能,也取决于它与新宿主蚊子基因组的相互作用。将使用Hermes转座元件来检验这一命题,Hermes转座元件是HAT转座元件家族的成员,也是埃及伊蚊的有效转化载体。这项建议的具体目的是:1)纯化和鉴定Hermes转座酶,2)了解Hermes元件的结构:功能关系,3)研究Hermes转座酶的相互作用,4)分离Hermes转座酶的高活性突变体,并测试它们在蚊子中的高活性。纯化的具有生物活性的Hermes转座酶将用于确定Hermes元件切除和转座的最佳物理和生物学参数,以及宿主因素在蚊子Hermes元件转座中可能发挥的作用。转座酶和元件的区域,如倒置末端重复序列和相邻的保守基序,是转座所需的,将被识别出来,并利用突变技术,确定它们在这一过程中的确切作用。我们已经确定在含有自主形式的Hermes元件的转基因昆虫中存在的Hermes元件的环形形式的作用将被调查。最后,真核和基于原核的表达系统都可以产生活性的Hermes转座酶,这两种系统都将被用于识别Hermes元件系统的超活性形式,然后将测试其在Ae中的活性升高水平。埃及伊蚊。
到目前为止,我们对用于遗传转化蚊子的转座元件在蚊子基因组中的作用或行为知之甚少。这现在构成了将基于转座子的技术扩展到具有重要医学意义的蚊子物种的最大瓶颈。这项技术将补充控制蚊媒疾病的现有方法,并将提供新的途径来处理因这些影响人类健康和国家经济的破坏性疾病的地理分布增加而产生的紧迫问题。这项新技术将使病原体传播所需的蚊子基因得以识别和分离,将提高鉴定转基因蚊子菌株的效率,并将提供有关防止病原体传播的基因通过转座元件在蚊子种群中传播的可能性的重要数据。
项目成果
期刊论文数量(0)
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PETER William ATKINSON其他文献
PETER William ATKINSON的其他文献
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{{ truncateString('PETER William ATKINSON', 18)}}的其他基金
Request for support for the Second International Symposium: Facing the Challenges
请求支持第二届国际研讨会:面对挑战
- 批准号:
8205403 - 财政年份:2012
- 资助金额:
$ 42.87万 - 项目类别:
Small RNA regulation of transponsons in mosquitoes.
蚊子转座子的小 RNA 调控。
- 批准号:
8084387 - 财政年份:2010
- 资助金额:
$ 42.87万 - 项目类别:














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