Isolation of Promoters Specifically Utilized in a Mosquito Host by a Bacterial Symbiont

Dawn C. Bisi

Defense Date

11-16-2005

Graduation Date

Fall 1-1-2005

Availability

Campus Only

Submission Type

thesis

Degree Name

MS

Department

Biological Sciences

Committee Chair

David J. Lampe

Committee Member

Joseph R. McCormick

Committee Member

Nancy Trun

Keywords

IVET, malaria, paratransgenesis

Abstract

Malaria is caused by the transmission of the parasite Plasmodium to humans by female Anopheles mosquitoes. This disease kills 1-2 million people annually and the number of deaths is increasing partly because of the failure of current control strategies. The effects of insecticides and drug treatments are muted by eventual evolution of resistances for both the insect vector and parasite. Recent genetic approaches to combating malaria are being developed. Bacteria that reside in the mosquito gut can be engineered to express antimalarial "effector" gene products that will inhibit Plasmodium development. The mosquito endosymbiont Enterobacter agglomerans was chosen as a potential candidate for carrying these transgenes in a wild mosquito population. Preliminary work towards this goal is presented here. IVET (in vivo expression technology) techniques were used to trap E. agglomerans promoter sequences and to select for those that are expressed under certain conditions within the mosquito and during conditions mimicking a blood meal. Promoters of this class can ensure effective expression of antimalarial molecules at the proper time and place inside the insect. The work presented here uncovered twenty-one E. agglomerans promoter and partial gene sequences identified via homology to genes of related bacterial species. Promoters of varying strengths as measured during growth on specific indicator media and in a colorimetric assay will be highlighted.

Format

PDF

Language

English

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