Using P-element Induced Male Recombination to Generate a Deletion in the DMAP1 Gene on Chromosome Two in Drosophila melanogaster
Abstract: The goal of this study was to induce a deletion in the DMAP1 gene on chromosome two in Drosophila melanogaster through P-element mobilization. The DMAP1 gene may be an essential gene, however not much is known about it. We attempted to uncover the function of DMAP1 by creating a series of genetic crosses and selecting for brown-eyed non-stubble male flies that may have the deletion. To test whether these flies had the deletion, we produced PCR products and ran them on an agarose gel, which resulted as inconclusive. We created a balanced stock of flies homozygous for the deletion to see if the
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Also, we made use of cinnabar (cn) and brown (bw) markers on either side of the P-element to select for chromosomes that may have deletions to one side or the other of the element when transposition occurs. Lastly, by using progeny that may have a DMAP1 deletion and balancer chromosomes, which contain dominant visible mutations so we can identify flies heterozygous for the balancer and contain recessive lethal mutations so that balancer/balancer flies do not survive, allowed us to create a balanced stock for the recombinant male chromosome with the possible deletions and therefore study its function. After selecting for flies that may have deletions in the DMAP1 gene, we used PCR analysis (polymerase chain reaction) to see if deletions have actually been made into DMAP1. We used these approaches because they allowed us to select for possible DMAP1 deletion events, they allowed us to identify which flies carried the recombinant chromosome with a possible DMAP1 deletion and which didn’t, and allowed us to have the ability to generate homozygous recombinant flies whenever we want for further study.
Materials and Methods: My group performed several crosses of flies in hopes of generating a deletion in the DMAP1 gene in F2 recombinant male flies. We made crosses by using CO2 gas and plexiglass platforms to