Development of the Bi-Partite Gal4-UAS System in the African Malaria Mosquito, Anopheles gambiae

Lynd, Amy ORCID: https://orcid.org/0000-0001-6054-0525 and Lycett, Gareth ORCID: https://orcid.org/0000-0002-2422-053X (2012) 'Development of the Bi-Partite Gal4-UAS System in the African Malaria Mosquito, Anopheles gambiae'. PLoS ONE, Vol 7, Issue 2, e31552.

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Abstract

Functional genetic analysis in Anopheles gambiae would be greatly improved by the development of a binary expression system, which would allow the more rapid and flexible characterisation of genes influencing disease transmission, including those involved in insecticide resistance, parasite interaction, host and mate seeking behaviour. The Gal4-UAS system, widely used in Drosophila melanogaster functional genetics, has been significantly modified to achieve robust application in several different species. Towards this end, previous work generated a series of modified Gal4 constructs that were up to 20 fold more active than the native gene in An. gambiae cells. To examine the Gal4-UAS system in vivo, transgenic An. gambiae driver lines carrying a modified Gal4 gene under the control of the carboxypeptidase promoter, and responder lines carrying UAS regulated luciferase and eYFP reporter genes have been created. Crossing of the Gal4 and UAS lines resulted in progeny that expressed both reporters in the expected midgut specific pattern. Although there was minor variation in reporter gene activity between the different crosses examined, the tissue specific expression pattern was consistent regardless of the genomic location of the transgene cassettes. The results show that the modified Gal4-UAS system can be used to successfully activate expression of transgenes in a robust and tissue specific manner in Anopheles gambiae. The midgut driver and dual reporter responder constructs are the first to be developed and tested successfully in transgenic An. gambiae and provide the basis for further advancement of the system in this and other insect species.

Item Type:	Article
Subjects:	QU Biochemistry > Genetics > QU 475 Genetic processes QX Parasitology > Insects. Other Parasites > QX 515 Anopheles QX Parasitology > Insects. Other Parasites > QX 600 Insect control. Tick control QX Parasitology > Insects. Other Parasites > QX 650 Insect vectors WA Public Health > Preventive Medicine > WA 110 Prevention and control of communicable diseases. Transmission of infectious diseases
Digital Object Identifer (DOI):	https://doi.org/10.1371/journal.pone.0031552
Depositing User:	Users 183 not found.
Date Deposited:	24 Feb 2012 16:20
Last Modified:	22 Aug 2019 12:14
URI:	https://archive.lstmed.ac.uk/id/eprint/2488

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