Robust gut-specific gene expression in transgenic Aedes aegypti mosquitoes
Robust gut-specific gene expression in transgenic Aedes aegypti mosquitoes
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DOI：
10.1073/pnas.97.20.10895
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发表时间：
2000-09-26
影响因子：
11.1
通讯作者：
Jacobs-Lorena, M
中科院分区：
文献类型：
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作者：
Moreira, LA;Edwards, MJ;Jacobs-Lorena, M
对人类疾病的蚊子载体的载体能力进行遗传修饰需要能够以适当的组织和阶段特异性驱动基因表达的启动子。我们报告了转基因埃及伊蚊中两种蚊子肠道特异性启动子的表征。邻近 Ae 编码区 5' 端的 1.4 kb DNA 片段。将埃及伊蚊羧肽酶 (AeCP) 基因和冈比亚按蚊羧肽酶 (AgCP) 基因 5' 端相应的 3.4-kb DNA 片段与萤火虫荧光素酶报告基因连接并引入到埃及伊蚊中。使用 Hermes 和 Mariner (Mos1) 转座子获得埃及伊蚊种系。使用 AeCP 构建体获得了 6 个独立的转基因系，1 个使用 AgCP 构建体。荧光素酶 mRNA 和蛋白质在 6 个 AeCP 系中的 4 个以及 AgCP 系的转基因蚊子的肠道中大量表达。报告基因的表达具有肠道特异性，并在血液摄入后约 24 小时达到峰值水平。 AeCP 和 AgCP 启动子可用于驱动阻碍蚊子肠道内寄生虫发育的基因表达。
Genetic modification of the vectorial capacity of mosquito vectors of human disease requires promoters capable of driving gene expression with appropriate tissue and stage specificity. We report on the characterization in transgenic Aedes aegypti of two mosquito gut-specific promoters. A 1.4-kb DNA fragment adjacent to the 5' end of the coding region of the Ae. aegypti carboxypeptidase (AeCP) gene and a corresponding 3.4-kb DNA fragment at the 5' end of the Anopheles gambiae carboxypeptidase (AgCP) gene were linked to a firefly luciferase reporter gene and introduced into the Ae. aegypti germ line by using Hermes and mariner (Mos1) transposons. Six independent transgenic lines were obtained with the AeCP construct and one with the AgCP construct. Luciferase mRNA and protein were abundantly expressed in the guts of transgenic mosquitoes in four of the six AeCP lines and in the AgCP line. Expression of the reporter gene was gut-specific and reached peak levels at about 24 h post-blood ingestion. The AeCP and AgCP promoters can be used to drive the expression of genes that hinder parasite development in the mosquito gut.