Tools for Anopheles gambiae Transgenesis.
Tools for Anopheles gambiae Transgenesis.
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DOI：
10.1534/g3.115.016808
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发表时间：
2015-04-13
期刊：
影响因子：
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通讯作者：
Marois E
中科院分区：
文献类型：
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作者：
Volohonsky G;Terenzi O;Soichot J;Naujoks DA;Nolan T;Windbichler N;Kapps D;Smidler AL;Vittu A;Costa G;Steinert S;Levashina EA;Blandin SA;Marois E
转基因是研究基因功能和在实验室生物体中引入所需特征的基本工具。由于生物生命特征的多样性和基因组信息方面的知识差距，在非模式生物中建立转基因是具有挑战性的。有些程序将广泛适用于许多生物体，而另一些程序则必须专门针对目标物种制定。自本世纪头十年以来，病媒蚊子中的转基因一直存在，但一直受到这些昆虫微妙生物学的限制。在这里，我们报道了我们为疟疾媒介冈比亚按蚊设计的转基因工具的汇编，包括新的对接菌株、方便的转基因质粒、扑霉素抗性选择标记、表达cre重组酶的蚊子以及定义克隆启动子活性的各种报告系。这个工具箱有助于在这个物种中实现转基因的例行公事，现在正在使日益精细的遗传操作的发展成为可能，例如靶向突变。这里报告的一些试剂和方法很容易转移到其他非模式物种，包括其他病媒或农业害虫。
Transgenesis is an essential tool to investigate gene function and to introduce desired characters in laboratory organisms. Setting-up transgenesis in non-model organisms is challenging due to the diversity of biological life traits and due to knowledge gaps in genomic information. Some procedures will be broadly applicable to many organisms, and others have to be specifically developed for the target species. Transgenesis in disease vector mosquitoes has existed since the 2000s but has remained limited by the delicate biology of these insects. Here, we report a compilation of the transgenesis tools that we have designed for the malaria vector Anopheles gambiae, including new docking strains, convenient transgenesis plasmids, a puromycin resistance selection marker, mosquitoes expressing cre recombinase, and various reporter lines defining the activity of cloned promoters. This toolbox contributed to rendering transgenesis routine in this species and is now enabling the development of increasingly refined genetic manipulations such as targeted mutagenesis. Some of the reagents and procedures reported here are easily transferable to other nonmodel species, including other disease vector or agricultural pest insects.