Microparticle Bombardment as a Method for Transgenesis in Auanema and Tokorhabditis.

Abstract

Functional gene analysis tools in Caenorhabditis elegans are often ineffective in other nematodes due to differences in gonadal morphology and transgene silencing. Here, we established a method to generate stable transgenic lines in the nematodes Auanema freiburgense and Tokorhabditis tufae using microparticle bombardment coupled with hygromycin B selection. Despite using non-codon-optimized GFP, transgenic strains expressing fluorescent markers were obtained in both species. Additionally, an Auanema codon-optimized RFP construct showed robust expression in all tissues. This method will be valuable for future studies into the unusual sex determination, viviparity, and stress resistance in Auanema and Tokorhabditis .