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Identifying housekeeping gene promoters in deer tick, Ixodes scapularis
Ixodes scapularis is a major vector of the bacterium, Borrelia burgdorferi, the main pathogen responsible for Lyme Disease. However, understanding of the acquisition and spread of tick-borne diseases has been hampered by the lack of transgenic tools. These tools include fluorescent markers under the control of housekeeping gene promoters as visual markers for successful genetic transformation. The development of constructs with I. scapularis-specific gene promoters will also be useful for expression of pathogen-targeting gene products. More specifically, once these promoters are developed and confirmed, we can utilize these promoters to express RNAi to knockdown genes of interest for vector capacity and pathogen acquisition. Additionally, promoters can be used to drive recombinant protein expression, including fluorescent gene expression. Developing fluorescent reporters will be useful as markers of successful genetic transformation. This will enable fast sorting of transgenic individuals by fluorescence while avoiding the need for regular DNA sequencing of individual progeny. During this project, I intend on identifying promoters of housekeeping genes and validating them via green fluorescent protein reporter assay in an Ixodes scapularis cell line.