UNIVERSITY OF MASSACHUSETTS
Our overall goal is to identify transcription factors (TFs) that can bind C. elegans gene promoters. This will further our understanding of the networks that control gene expression and to delineate the regulatory genomic code. In the first year of the Project, we have further optimized our yeast one-hybrid (Y1H) pipeline. This resulted in a more convenient assay system and higher fidelity output. However, we have also encountered two bottlenecks, mostly regarding bait strain generation and plate pouring capacity. It is the goal of this request to alleviate these bottlenecks to ensure continued progress on the Project, at a high tempo and, of course, with high fidelity. Briefly, we requested funds to ensure that we can generate the DNA bait strains in a timely manner, that we can provide the project with sufficient plates/media, and finally to clone TFs that we currently do not yet have in our collection; we have 800 of the 940 TFs available, and new gene models are available for ~55 that are missing. This will enable us to screen 48 baits per week, rather than 24 (our current throughput), and thereby to achieve the goals that we outlined in the original proposal.