| | Category | MI | L29 | Isolated Meiosis Related Genes in Fungi |
| | Abstract | Isolating Genes Related to Meiosis in Fungi |
| | |
| | The genes Rad51 and Dmc1 are genes known to exist in virtually all |
| | organisms that carry out meiosis. Rad51 is present in both mitosis and |
| | meiosis, while Dmc1 is a meiosis specific gene. Confirmation of its |
| | presence and sequencing of these genes within various species of fungi |
| | (ascomycetes and basidiomycetes) allows for several comparisons that |
| | can give insight on the relation of species to one another, or the nature in |
| | which meiosis has possibly evolved. The loss or degeneracy of either |
| | gene can suggest that meiosis within a species has been lost, or has |
| | evolved in a way that no longer requires Rad51/Dmc1. The project’s |
| | objective was to confirm the presence or lack of Rad51/Dmc1 within |
| | various species of fungi. |
| | |
| | Samples are run through a series of PCR reactions and gel electrophoresis |
| | in order to determine the size potential Rad51 or DMC1 gene fragments. |
| | Several “nested” PCRs may be run until a viable band of DNA is found. |
| | Once bands are found, the sample is run through a low melt gel in order to |
| | isolate the gene fragment for cloning. The genes are cloned via E. coli |
| | plasmid cloning vectors and subject to further PCR screening in order to |
| | confirm the existence of the targeted gene fragment of the correct size. If |
| | correct, the colonies of the correct insert are cultured then purified to |
| | isolate the desired plasmid DNA, which is sent in to be sequenced. |
| | |
| | The project identified at total of 18 species with Rad51, and 4 species with |
| | Dmc1. Several fragments of Rad51 were successfully extended as well. |
| | The project identified several samples of fungi DNA with contaminations. |
| | Confirmations of Rad51 and Dmc1 within species of fungi with no previous |
| | genotypes were also found. Bisporella sp. was found to have two |
| | varying copies of Rad51, suggesting possible gene duplication. |
| | Bibliography | 1) Schurko, A.M. and J. M. Logsdon, Jr. 2008. Using a meiosis detection |
| | toolkit to investigate ancient asexual scandals and the evolution of sex. |
| | BioEssays, 30:6:579 - 589. 2) Villeneuve, Anne M., and Kenneth J. Hillers. |
| | "Whence Meiosis?" Cell. Vol. 106. Cell, 2001. 647-50. Print. 3) Birren, |
| | Bruce W. "The Fungal Genome Initiative and Lessons Learned from |
| | Genome Sequencing." Methods in Enzymology. By Christina A. Cuomo. |
| | Vol. 470. Elsevier, 2010. 834-53. Print. |