Isolation of deletion mutants by reverse genetics incaenorhabditis elegans

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Abstract
Obtaining mutant animals is important for studying the function of a particular gene. A chemical mutagenesis was first carried out to generate mutations in C. elegans. In this study, we used ultraviolet‐activated 4,5’,8‐trimethylpsoralen to induce small deletion mutations. A library of muta‐genized worms was prepared for recovery of candidate animals and stored at 15°C during screening instead of being made into a frozen stock library. In order to isolate deletion mutations in target genes, a polymerase chain reaction (PCR)‐based screening method was used. As a result, two independent mutants with deletions of approximately 1.0 kb and 1.3 kb were isolated. This modified and improved reverse genetic approach was proven to be effective and practical for isolating mutant animals to study gene function at the organismal level.