Researcher(s)
- Benjamin Sekowski, Biological Sciences, University of Delaware
Faculty Mentor(s)
- Amber Krauchunas, , University of Delaware
Abstract
The goal of my research is to find if the proteins COPA-1 and SNAP-1 are important for fertility. I created two strains that each express TIR1 in the germline and the degron-tagged protein of interest, either copa-1 or snap-1. Males carrying the TIR1 transgene were crossed with the copa-1 and snap-1 hermaphrodites. PCR was used to identify which worms were homozygous for both TIR1 and copa-1 and homozygous for both TIR1and snap-1. The two resulting lines that were created were wrdSi51 (germline TIR1); copa-1::gfp::AID and wrdSi51 (germline TIR1); snap-1::gfp::AID. The next step was to find out how TIR1-mediated degradation of the protein of interest will affect reproduction. Broodsizing was conducted to measure the number of progeny produced when the targeted protein COPA-1 is degraded in the germline. Broodsizing also occurred on a control group of N2 wild type worms. The results of the brood counting were that the N2 wild type control group reproduced normally, while the line with degraded COPA-1 protein did not reproduce at all. No embryos or eggs were seen on the plates. The proteins COPA-1 and SNAP-1 were also tagged with GFP to observe where they are expressed. The worms with the desired homozygous germline were imaged with the widefield fluorescence microscope. The COPA-1::GFP expression was observed to be consistent in the spermatheca. In conclusion, the COPA-1 protein does have an effect on reproduction and is expressed in the spermatheca.