Analysis of apyrase 5′ upstream region validates improved Anopheles gambiae transformation technique

被引：18

作者：

Lombardo F. ^{[1
,4
]}

Lycett G.J. ^{[2
,5
]}

Lanfrancotti A. ^{[1
,6
]}

Coluzzi M. ^{[1
]}

Arcà B. ^{[1
,3
]}

机构：

[1] Department of Public Health, Parasitology Section, Sapienza University of Rome, 00185, Roma

[2] European Molecular Biology Laboratory, 69117-Heidelberg

[3] Department of Structural and Functional Biology, University Federico II, 80126, Napoli, Via Cinthia

[4] Imperial College London, SW7 2AZ, London, Imperial College Road

[5] Liverpool School of Tropical Medicine, L3 5QA, Liverpool, Pembroke Place

[6] Kennedy Institute of Rheumatology, Imperial College London, W6 8LH, London

来源：

BMC Research Notes | / 2卷 / 1期

基金：

英国生物技术与生命科学研究理事会;

关键词：

Salivary Gland; Hemocyte; Salivary Gland Extract; Mosquito Anopheles Gambiae; Efficient Genetic Transformation;

D O I：

10.1186/1756-0500-2-24

中图分类号：

学科分类号：

摘要：

Background. Genetic transformation of the malaria mosquito Anopheles gambiae has been successfully achieved in recent years, and represents a potentially powerful tool for researchers. Tissue-, stage- and sex-specific promoters are essential requirements to support the development of new applications for the transformation technique and potential malaria control strategies. During the Plasmodium lifecycle in the invertebrate host, four major mosquito cell types are involved in interactions with the parasite: hemocytes and fat body cells, which provide humoral and cellular components of the innate immune response, midgut and salivary glands representing the epithelial barriers traversed by the parasite during its lifecycle in the mosquito. Findings. We have analyzed the upstream regulatory sequence of the An. gambiae salivary gland-specific apyrase (AgApy) gene in transgenic An. gambiae using a piggyBac transposable element vector marked by a 3xP3 promoter:DsRed gene fusion. Efficient germ-line transformation in An. gambiae mosquitoes was obtained and several integration events in at least three different G0 families were detected. LacZ reporter gene expression was analyzed in three transgenic lines/groups, and in only one group was tissue-specific expression restricted to salivary glands. Conclusion. Our data describe an efficient genetic transformation of An. gambiae embryos. However, expression from the selected region of the AgApy promoter is weak and position effects may mask tissue- and stage- specific activity in transgenic mosquitoes. © 2009 Lombardo et al.

引用

共 22 条

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