Transcript Drosophila Melanogaster

Molecular Characterization of an
Unknown P-Element Insertion in
Drosophila melanogaster
Why Drosophila melanogaster?

Benefits of Drosophila as a model
organism:
1.
2.
3.
4.
5.
6.
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
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”
P{lacW}
white gene
 Plasmid pBR322
 6 restriction enzyme sites

Plasmid Rescue

An inserted
plasmid is
recovered from
genome of
Drosophila to
clone the plasmid
along with
adjacent genomic
sequences
pBR322
The Four Steps of Plasmid Rescue
http://upload.wikimedia.org/wikipedia/en/f/fa/P_elements_2.png
The Four Steps (Continued)
Results
Expected
Observed
Controls
Control
Expected Observed
No DNA
No growth
No growth
pBR322
Growth
No growth
pBR322 + EcoRI
+ligase
Growth
No growth
pBR322 + EcoRI
No growth
No growth
Discussion

Why didn’t it work?
Genomic DNA purification failed
 Ligation failed
 Plasmid degraded
pBR322
No growth
 Failure in making the plates

Bacteria+
killed
in plating
pBR322
EcoRI
No growth
 E. coli
not
competent
+ligase

pBR322
No growth
References

Cook, HA., ed. Molecular Characterization of an Unknown Pelement 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
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http://neofronteras.com/wp-content/photos/ drosophila.jpg

http://www.anselm.edu/homepage/jpitocch/genbio/transposons.J
PG
Acknowledgements
We would like to thank Dr.Cook, Kirsten, and
Dr.Miyamoto for their guidance, support,
and overall awesomeness!!
We love you!!!
The
End