The Search for Active Transposable Elements in the Aedes aegypti Genome
Mentor:James Burnette, Academic Coordinator, University of California, Riverside
Aedes aegypti is a mosquito vector for human pathogens such as yellow fever, dengue fever, and malaria. Transposable elements (TEs) are short sequences of DNA that can move to different places in the genome. DNA TEs are commonly used as genetic tools to get a better understanding of an organism's genome. However, exogenous TEs do not function in A. aegypti. The purpose of this research was to identify putatively active transposable elements endogenous to A. aegypti by bioinformatics, isolate the candidates, and verify their activity experimentally for development as genetic tools in A. aegypti. Several techniques such as PCR, gel electrophoresis, and cloning can be used to determine TEs activity. Potentially active TEs were identified using bioinformatics. A bioinformatics program was used to predict the DNA sequences of putatively active TEs. After extracting DNA from A. aegypti mosquitoes, PCR was used to amplify and confirm its presence. We analyzed the DNA using gel electrophoresis to see if the size of the sequence matched the one predicted by bioinformatics. These TEs were introduced into plasmids and cloned into E. coli. Plasmid PCR was performed and the DNA was sent for sequencing to confirm the presence of the TE. In order to test for their activity, these TEs will need to be cloned into yeast. If the TE is active in the yeast assay, it will be tested in mosquitos. Any TE active in yeast is a candidate for genetic manipulation of the mosquito genome. We attempted to clone the TEs, but the sequence results were not clean enough to be analyzed.