Introducing Fluorescent Fusion Constructs Into C. elegans By Microparticle Bombardment

Proper chromosome segregation during meiosis depends on synapsis and recombination of homologous chromosomes in Prophase I. Because improper meiotic segregation can result in aneuploidy and contribute to infertility and birth defects, we are interested in learning what mechanisms ensure that synapsis and recombination take place correctly. We are studyingseveral proteins, encoded in the genome of C. elegans that may regulate these events. To gain insight into their functions, we want to localize these proteins in the germlines of live animals. To do this we have used Multisite Gateway technology to generate fusion constructs of each gene with GFP and mCherry, fluorescent tags that enable us to visualize the proteins in vivo. The constructs are under the control of the germline specific pie-1 promoter and include their endogenous 3'UTR. They will be introduced into C. elegans using microparticle bombardment with the unc-119 gene as a selective marker. Our expected results are transformants that will exhibit wild-type phenotype and express the fluorescent proteins during meiosis. Our progress in generating these transgenic animals will be presented and discussed.