Generation of Transgenic Caenorhabditis Elegans by Multiple Transformation Methods
by Shane Clune, Arinda Abbuhl, and Elizabeth Ahrendsen
Faculty advisor: Dr. James Hampton
The purpose of our research project is to effectively transform the nematode C. elegans via three different transformation techniques: Aerosol Beam Injection, microparticle bombardment, and a Particle Infusion Gun (PIG). A comparison of these three methods may provide a more effective way to transform C. elegans, a useful genetic model organism.
To test the effectiveness of the three methods in nematodes, we attempted to resinsert a gene into a knockout strain of C. elegans called unc-119. The unc-119 gene affects a collagen protein of C. elegans, causing the mutant nematodes to move in an uncoordinated manner and prevents their larvae from entering into a suspended animation state known as dauer. To rescue the phenotype we inserted a plasmid containing the unc-119 gene. This plasmid also provided us with an internal control since it will also impart ampicillin resistance to E. coli.
Different parameters were tested; including time each section of the plate was subjected to bombardment and concentration of TRIS buffer used to suspend the DNA. Other things to be considered are age of the worms being bombarded and care of the worms after we attempted to insert the plasmid. We will present the results of the research, as well as outline future studies.