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

2.  Benefits of Drosophila as a model organism: 1. Short generation time 2. Small, easy to care for 3. Large number of offspring 4. Easy to see many heritable traits 5. Eight chromosomes 6. Complete genome has been sequenced and has an estimated 13,600 genes Why Drosophila melanogaster?

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”

5. P{lacW}  white gene  Plasmid pBR322  6 restriction enzyme sites

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

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

8. The Four Steps (Continued)

9. Results Expected Observed

10. Controls ControlExpectedObserved No DNANo growth pBR322GrowthNo growth pBR322 + EcoRI +ligase GrowthNo growth pBR322 + EcoRINo growth

11. Discussion  Why didn’t it work? Genomic DNA purification failed Ligation failed pBR322No growth pBR322 + EcoRI +ligase No growth Plasmid degraded Failure in making the plates Bacteria killed in plating pBR322No growth E. coli not competent

12. 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.ht m  http://neofronteras.com/wp-content/photos/ drosophila.jpg  http://www.anselm.edu/homepage/jpitocch/genbio/transposons.J PG

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

14. The End