HTSS assay

The HTSS is used to test the effects of new chemicals on the behavior of adult mosquitoes, including contact irritancy, spatial repellency, and toxic actions. The modular device (Figure 6.3) uses different arrays of aluminum (test) and Plexiglas (control) cylinders, depending on the objective of the assay. 

While S. pombe strains carrying mutations that lower PKA activity can be detected by their ability to form colonies on solid medium lacking uracil [2], this phenotype is not sufficiently robust for HTSs to detect compounds that reduce PKA activity. Recently, we constructed zn. fbpl-driven GFP reporter that allows for small-molecule screens to detect compounds that stimulate PDF activity or inhibit AC (or their associated Gas) or PKA proteins [9]. Cell-based assays utilizing the fbpl-um4 and/ pi-GFP reporters allow for inexpensive HTSs for small-molecule modulators of cloned PKA pathway genes expressed in S. pombe. 

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