Robust heat-inducible gene expression by two endogenous hsp70-derived promoters in transgenic Aedes aegypti.
Robust heat-inducible gene expression by two endogenous hsp70-derived promoters in transgenic Aedes aegypti.
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DOI：
10.1111/j.1365-2583.2011.01116.x
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发表时间：
2012-02
影响因子：
2.6
通讯作者：
Adelman ZN
中科院分区：
文献类型：
--
作者：
Carpenetti TL;Aryan A;Myles KM;Adelman ZN
埃及伊蚊是引起登革热、登革出血热和黄热病的病毒的重要媒介。研究这种蚊子基因功能的反向遗传方法受到缺乏强有力的诱导型启动子的限制，无法精确控制蛋白质编码或发夹RNA转基因。同样，研究载体感受态的分子和生物化学基础将受益于在感染期间的特定时间激活抗病原体分子的能力。我们已经表征了来自两个Ae的基因组序列的能力。埃及热休克蛋白70基因，以驱动瞬时和种系转化环境中报告基因的热诱导表达。AaHsp 70-荧光素酶转录本在热休克后特异性积累，并显示出与内源性AaHsp 70基因相似的快速诱导和衰变模式。转基因Ae.在热休克后4小时，埃及伊蚊在整个成虫中增加了10.25 -50倍，在24小时仍有显著的活性（10.20倍）。所描述的AaHsp 70启动子对于基因功能研究以及抗病原体分子表达的精确定时可能是有价值的。
Aedes aegypti is an important vector of the viruses that cause dengue fever, dengue hemorrhagic fever, and yellow fever. Reverse genetic approaches to the study of gene function in this mosquito have been limited by the lack of a robust inducible promoter to allow precise temporal control over a protein-encoding or hairpin RNA transgene. Likewise, investigations into the molecular and biochemical basis of vector competence would benefit from the ability to activate an anti-pathogen molecule at specific times during infection. We have characterized the ability of genomic sequences derived from two Ae. aegypti hsp70 genes to drive heat-inducible expression of a reporter in both transient and germline transformation contexts. AaHsp70-luciferase transcripts accumulated specifically after heat shock, and displayed a pattern of rapid induction and decay similar to endogenous AaHsp70 genes. Luciferase expression in transgenic Ae. aegypti increased by ∼25-50 fold in whole adults by four hours after heat-shock, with significant activity (∼20 fold) remaining at 24 hr. Heat-induced expression was even more dramatic in midgut tissues, with one strain showing a ∼2500-fold increase in luciferase activity. The AaHsp70 promoters described could be valuable for gene function studies as well as for the precise timing of the expression of anti-pathogen molecules.
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影响因子：
6.7
作者：
Isaacs AT;Li F;Jasinskiene N;Chen X;Nirmala X;Marinotti O;Vinetz JM;James AA
通讯作者：
James AA
影响因子：
2.6
作者：
Mathur, G.;Sanchez-Vargas, I.;James, A. A.
通讯作者：
James, A. A.
DOI：
10.1073/pnas.1000251107
发表时间：
2010-03-09
影响因子：
11.1
作者：
Fu, Guoliang;Lees, Rosemary S.;Alphey, Luke
通讯作者：
Alphey, Luke
DOI：
10.1073/pnas.0701515104
发表时间：
2007-06-12
影响因子：
11.1
作者：
Adelman, Zach N.;Jasinskiene, Nijole;James, Anthony A.
通讯作者：
James, Anthony A.
DOI：
10.1073/pnas.97.20.10895
发表时间：
2000-09-26
影响因子：
11.1
作者：
Moreira, LA;Edwards, MJ;Jacobs-Lorena, M
通讯作者：
Jacobs-Lorena, M