Gene knockout by inducing P-element transposition in Drosophila

Abstract

The P-element has been successfully used in germline transformation to create transgenic flies and as an insertion mutagenesis agent to disrupt genes. Moreover, P-element can also be used to knockout genes that are near the insertion sites by inducing its imprecise transposition. In this article, P-element insertion lines were crossed with transposase expression fly to recover the transposon, and the deletion of flanking sequence, which was created by imprecise excision, was detected by PCR. Through this method, null alleles of seven genes that spread over three major chromosomes (X, second, third) were generated in Drosophila melanogaster. Results show that the frequency of flanking deletions is expected to be 1%, and it is enough to pick up at least one ideal deletion line from 200 independent recovery lines in general. These data suggest that although the frequency of disrupted gene varied greatly, from 0.13 to 2.34%, gene knockout by inducing P-element transposition appears to be a feasible and effective strategy, compared to the complicated process of gene targeting based on homologous recombination.