Construction of a C. elegans Strain Lacking Biogenic Amine Neurotransmitters

Student Presenter(s): Dianna Levin, Kevin Lee
Faculty Mentor: Navin Pokala
Department: Life Sciences
School/College: College of Arts and Sciences, Long Island

Caenorhabditis elegans is a useful model organism for neuroscience because of its fully mapped brain wiring, and well characterized genome. One gene of interest is the cat-1 gene, which is responsible for the transport of serotonin, dopamine, octopamine and tyramine into synaptic vesicles. These neurotransmitters are crucial for the function of laying eggs, pharyngeal pumping, locomotion and learning. A mutant strain that contains a deletion in cat-1 has been generated. Unfortunately, due to the way this deletion was generated (UV radiation), this mutant strains contain additional unwanted mutations. To get rid of these unwanted mutations, only leaving the mutation ib the cat-1 gene, the process of backcrossing was implemented. Mutant cat-1 hermaphrodites were crossed with wild-type (normal) males and their male offsprings were picked. The male offspring were crossed with wild-type hermaphrodites. Each of these crosses replaces mutant DNA with wild-type DNA. Offspring that retain for the cat-1 mutation were identified using PCR. By doing rounds of this process, unwanted mutations can be diluted out.