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Molecular Characterization of an Unknown P-Element Insertion in Drosophila melanogaster

Molecular Characterization of an Unknown P-Element Insertion in Drosophila melanogaster. Why Drosophila melanogaster ?. Benefits of Drosophila as a model organism: Short generation time Small, easy to care for Large number of offspring Easy to see many heritable traits

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Molecular Characterization of an Unknown P-Element Insertion in Drosophila melanogaster

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  1. Molecular Characterization of an Unknown P-Element Insertion in Drosophila melanogaster

  2. Why Drosophila melanogaster? • Benefits of Drosophila as a model organism: • Short generation time • Small, easy to care for • Large number of offspring • Easy to see many heritable traits • Eight chromosomes • Complete genome has been sequenced and has an estimated 13,600 genes

  3. Transposon Mutagenesis • Method for inducing mutations using a “genetically engineered” transposon to study function of genes • Transposon - Sequence of DNA that can move around to different positions within the genome of a single cell in a process called transposition. • AKA “jumping gene” or “mobile genetic element”

  4. P{lacW} • white gene • Plasmid pBR322 • 6 restriction enzyme sites

  5. Plasmid Rescue • An inserted plasmid is recovered from genome of Drosophila to clone the plasmid along with adjacent genomic sequences pBR322

  6. The Four Steps of Plasmid Rescue http://upload.wikimedia.org/wikipedia/en/f/fa/P_elements_2.png

  7. The Four Steps (Continued)

  8. Results Expected Observed

  9. Controls

  10. Discussion • Why didn’t it work? • Genomic DNA purification failed • Ligation failed • Plasmid degraded • Failure in making the plates • Bacteria killed in plating • E. coli not competent

  11. References • Cook, HA., ed. Molecular Characterization of an Unknown P-element insertion in Drosophila Melanogaster. 2006. 28 p. • http://www.discoverygenomics.net/images/sbts_cut-paste.gif • http://cwx.prenhall.com/brock/chapter10/medialib/pbr322.jpg • http://www.emulimited.com/images/SwimmingPools/image016.jpg • http://www.uoguelph.ca/pdc/Factsheets/Insect/DrosophilaFiles.htm • http://neofronteras.com/wp-content/photos/ drosophila.jpg • http://www.anselm.edu/homepage/jpitocch/genbio/transposons.JPG

  12. Acknowledgements We would like to thank Dr.Cook, Kirsten, and Dr.Miyamoto for their guidance, support, and overall awesomeness!! We love you!!!

  13. The End

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