Hermes Element Transposition in Mosquitoes

蚊子中的赫耳墨斯元素转位

基本信息

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转基因蚊子中引入的转座因子的行为,转基因蚊子是人类疾病的载体。本文所述研究的中心命题是,转座因子在实验室中用作蚊子的有效遗传工具以及在田间用作有益效应基因的有效“基因驱动机制”的能力由转座因子如何发挥功能以及它如何与新的宿主蚊子基因组相互作用决定。将使用爱马仕转座因子检验该命题,该转座因子是转座因子的hAT家族的成员,并且是蚊子埃及伊蚊的有效转化载体。因此,该提案的具体目标是:1/纯化和表征爱马仕转座酶,2)了解爱马仕元件的结构:功能关系,3)检查爱马仕转座酶的相互作用,以及4)分离爱马仕转座酶的过度活跃突变体并测试它们在蚊子中的过度活跃。纯化的生物活性爱马仕转座酶将用于确定爱马仕元件切除和转座的最佳物理和生物学参数以及宿主因子在蚊子中爱马仕元件转座中可能发挥的作用。将鉴定转座所需的转座酶和元件的区域,如反向末端重复序列和相邻的保守基序,并使用诱变确定它们在该过程中的确切作用。将研究我们已经鉴定存在于含有自主形式的爱马仕元件的转基因昆虫中的爱马仕元件的环状形式的作用。最后,基于真核和原核的表达系统都产生活性爱马仕转座酶,将用于鉴定爱马仕元件系统的过度活性形式,然后测试Ae中活性水平的升高。埃及人。 到目前为止,我们对用于遗传转化蚊子的转座因子在蚊子基因组中的工作或行为知之甚少。这现在构成了将基于转座元件的技术扩展到医学上重要的蚊子物种的最重要的瓶颈。这项技术将补充现有的控制蚊媒疾病的方法,并将提供新的途径来解决这些破坏性疾病的地理分布增加所产生的紧迫问题,这些疾病影响人类健康和国民经济。这一新技术将使病原体传播所需的蚊子基因得以鉴定和分离,将提高鉴定转基因蚊子品系的效率,还将提供关于防止病原体传播的基因通过转座因子在蚊子种群中传播的可能性的重要数据。

项目成果

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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:
HERMES ELEMENT TRANSPOSITION IN MOSQUITOES
蚊子中的 Hermes 元素转座
  • 批准号:
    6374205
  • 财政年份:
    1999
  • 资助金额:
    $ 42.87万
  • 项目类别:
HERMES ELEMENT TRANSPOSITION IN MOSQUITOES
蚊子中的 Hermes 元素转座
  • 批准号:
    2887986
  • 财政年份:
    1999
  • 资助金额:
    $ 42.87万
  • 项目类别:
Hermes Element Transposition in Mosquitoes
蚊子中的赫耳墨斯元素转位
  • 批准号:
    7163725
  • 财政年份:
    1999
  • 资助金额:
    $ 42.87万
  • 项目类别:
Hermes Element Transposition in Mosquitoes
蚊子中的赫耳墨斯元素转位
  • 批准号:
    6724405
  • 财政年份:
    1999
  • 资助金额:
    $ 42.87万
  • 项目类别:
Hermes Element Transposition in Mosquitoes
蚊子中的赫耳墨斯元素转位
  • 批准号:
    7337296
  • 财政年份:
    1999
  • 资助金额:
    $ 42.87万
  • 项目类别:
HERMES ELEMENT TRANSPOSITION IN MOSQUITOES
蚊子中的 Hermes 元素转座
  • 批准号:
    6534169
  • 财政年份:
    1999
  • 资助金额:
    $ 42.87万
  • 项目类别:
Hermes Element Transposition in Mosquitoes
蚊子中的赫耳墨斯元素转位
  • 批准号:
    7010012
  • 财政年份:
    1999
  • 资助金额:
    $ 42.87万
  • 项目类别:
HERMES ELEMENT TRANSPOSITION IN MOSQUITOES
蚊子中的 Hermes 元素转座
  • 批准号:
    6170393
  • 财政年份:
    1999
  • 资助金额:
    $ 42.87万
  • 项目类别:
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