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Show 76 Tyler Shimko college of science The ability to insert and inactivate genes in model organisms is crucial to uncovering the function of genes. We work on the nematode C. elegans and are developing a unique method of inserting single-copy transgenes. Our new method of insertion required the creation of landing site within the genome. This required that we first create a plasmid that contained the landing site we wished to utilize. Included in the plasmid was the landing site, which included a transposable element flanked by two selection markers. The first selec-tion marker, coding for the rescue of unc-18, allowed selection of only worms that had incorporated the landing site, based upon their ability to move naturally. The second selection marker, coding for a green fluorescent protein in the pharynx, allowed us to visually track which worms contained the landing site through a series of outcrosses. Once we had strains that contained the landing site, we had to identify the position of the landing site within the genome. This was done through inverse PCR that amplified a stretch of DNA corresponding to part of the landing site and part of the adjacent genomic DNA. These PCR prod-ucts were then sent for sequencing and the genomic portions of the amplified DNA were aligned with known portions of the genome to identify the precise insertion sites. Our preliminary results show that three of the eleven strains have landing sites in usable regions of the genome. Several other strains incorporated two landing sites consecutively, and are therefore unusable. Next, we plan to insert transgenes into the landing sites and test and assess the expression of the trans-gene. We will also use these strains, each with the landing site in a different region of the genome, to begin to assess how the genomic environment affects expression levels. UNIVERSAL TRANSGENE INSERTION IN C. ELEGANS Tyler Shimko (Erik Jorgensen) Department of Biology University of Utah UNDERGRADUATE RESEARCH ABSTRACTS Erik Jorgensen |