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Optimization of the Gal4-UAS system in an Anopheles gambiae cell line

Lynd, Amy ORCID: and Lycett, Gareth ORCID: (2011) 'Optimization of the Gal4-UAS system in an Anopheles gambiae cell line'. Insect Molecular Biology, Vol 20, Issue 5, pp. 599-608.

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The development of the bipartite Gal4‐UAS system in Anopheles gambiae would improve the functional characterization of genes in this important malaria vector. Towards this aim, we used Gal4 driver plasmids to successfully activate expression of the reporter gene, luciferase, from UAS responder plasmids when cotransfected into an An. gambiae cell line. To optimize Gal4‐regulated gene expression in mosquitoes, we compared the efficiency of a series of alternative Gal4 transactivators to drive reporter gene expression from responder plasmids incorporating different numbers of tandemly arrayed Gal4 binding sites or upstream activation sequences (UAS). The results indicated that the native Gal4 is only weakly active in these cells. Modified forms of Gal4, including those carrying minimal VP16 activation domains, as well as a deleted form of Gal4, give up to 20‐fold greater activity than the native protein, when used in conjunction with a responder plasmid having 14 UAS repeats. The identification of Gal4‐UAS vectors that are efficiently expressed in a mosquito cell line should facilitate the transfer of this versatile expression system to An. gambiae, and potentially to other insects of medical importance.

Item Type: Article
Subjects: QU Biochemistry > Genetics > QU 470 Genetic structures
QX Parasitology > Insects. Other Parasites > QX 515 Anopheles
Digital Object Identifer (DOI):
Depositing User: Users 183 not found.
Date Deposited: 26 Sep 2011 08:56
Last Modified: 22 Aug 2019 12:14


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