Evaluating and Isolating Promoters in Impatiens walleriana: Towards the Development of Mosquitocidal Nectar
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Mosquito-borne diseases, such as malaria, are a persistent problem, leading to numerous deaths across the world that could be easily prevented by better control of mosquito populations. Because the main source of energy for many mosquitoes are the sugars present in nectar, a novel approach for mosquito control that would complement methods already in use would be to develop plants that express mosquito-specific toxins in their nectar. Nectar-specific promoters would be a necessary step in developing plants that express mosquito toxin solely in their nectar. One nectar-specific promoter that has been found and sequenced is the pNEC promoter of Nicotiana langsdorffii X N. sanderae. To test the effectiveness of this promoter in Impatiens walleriana, pNEC was combined with enhanced green fluorescent protein (EGFP) and used in the transformation of I. walleriana. Kanamycin resistance was used to select transformants; presence of EGFP in nectar would then prove successful promotion of expression by pNEC. When no EGFP was found to be present, further investigation determined that the transformations were not successful, despite surviving the kanamycin selection step. A means for potentially isolating the nectar-specific promoter endogenous in I. walleriana was also tested. Thermal asymmetric interlaced PCR (TAIL-PCR) was used to successfully isolate the 26S rRNA promoter of I. walleriana, verifying it as a method to eventually isolate the IW23 promoter of I. walleriana.