Using P-Element Induced Male Recombination to Generate a Deletion in the Dmap1 Gene on Chromosome Two in Drosophila Melanogaster
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 deletion affected an essential gene. In one balanced stock, the gene deleted was found to be homozygous lethal, which may or may not be due to deletion of DMAP1. Therefore, further work is needed to confirm that the P-element created a deletion into DMAP1 (and extent of deletion) and that the lethality associated with the deletion is entirely due to a deletion in only DMAP1. In the case that lethality is not associated with deletion of the DMAP1 gene, further work is needed to determine its function temporally and spatially.
Introduction:
The objective of this experiment was to use a pre-existing P-element insertion near the DMAP1 gene on chromosome 2 in Drosophila melanogaster (fruit fly) to create a deletion in it. The CG33785 gene is located to the left of the P-element and the DMAP1 gene is located to the right. In vertebrates, DMAP1 encodes proteins that are involved in DNA methylation of C residues in DNA, which is associated with transcriptional inactivation (Fitzpatrick, 2013a). The DMAP1 gene is of interest because not much is currently known about its role in flies, and could possibly be an essential gene, a gene required for regulating survival, fertility, and/or development. One current possibility is that DMAP1 in flies
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 deletion affected an essential gene. In one balanced stock, the gene deleted was found to be homozygous lethal, which may or may not be due to deletion of DMAP1. Therefore, further work is needed to confirm that the P-element created a deletion into DMAP1 (and extent of deletion) and that the lethality associated with the deletion is entirely due to a deletion in only DMAP1. In the case that lethality is not associated with deletion of the DMAP1 gene, further work is needed to determine its function temporally and spatially.
Introduction:
The objective of this experiment was to use a pre-existing P-element insertion near the DMAP1 gene on chromosome 2 in Drosophila melanogaster (fruit fly) to create a deletion in it. The CG33785 gene is located to the left of the P-element and the DMAP1 gene is located to the right. In vertebrates, DMAP1 encodes proteins that are involved in DNA methylation of C residues in DNA, which is associated with transcriptional inactivation (Fitzpatrick, 2013a). The DMAP1 gene is of interest because not much is currently known about its role in flies, and could possibly be an essential gene, a gene required for regulating survival, fertility, and/or development. One current possibility is that DMAP1 in flies
References: Fitzpatrick, K (2013a) Class Project Slides: https://webct.sfu.ca/webct/entryPageIns.dowebct (accessed Apr 12, 2013). Fitzpatrick, K (2013b) Lecture 3 Notes on Fly Stuff: https://webct.sfu.ca/webct/entryPageIns.dowebct (accessed Apr 12, 2013). Fitzpatrick, K (2013c) Lab 2 Handout on Drosophila melanogaster Genetics: https://webct.sfu.ca/webct/entryPageIns.dowebct (accessed Apr 12, 2013). Fitzpatrick, K (2013d) Lecture 5 Notes on P-elements: https://webct.sfu.ca/webct/entryPageIns.dowebct (accessed Apr 12, 2013). Fitzpatrick, K (2013e) Format for Lab Write-Ups Handout: https://webct.sfu.ca/webct/entryPageIns.dowebct (accessed Apr 12, 2013). Fitzpatrick, K (2013f) Lecture Notes on Male Recombination Project: https://webct.sfu.ca/webct/entryPageIns.dowebct (accessed Apr 12, 2013). Fitzpatrick, K (2013g) Lab Handout for Week of March 19: https://webct.sfu.ca/webct/entryPageIns.dowebct (accessed Apr 12, 2013). Fitzpatrick, K (2013h) Lab Handout for Week of April 2: https://webct.sfu.ca/webct/entryPageIns.dowebct (accessed Apr 12, 2013). Appendix 2: PCR Summary Figure 2: templates used for PCR analysis (Fitzpatrick, 2013f) c) Heterozygous wildtype-eyed flies collected from this year (progeny from F2 cross in figure 1); 22-13-bw is the recombinant chromosome (top) and SM6a is a balancer (bottom) Figure 3: map of primer positions (Fitzpatrick, 2013f)