Transcript Presentation1 (Powerpoint)

Mouse Models of Prostate Cancer
Name: Vicki Javier
Mentor: Todd Hricik
Institution: MSKCC
Prostate Cancer
• Prostate Cancer: disease marked by
abnormal growth of tumor cells in the
prostate gland
• Prostate Cancer is detected by
– Digital Rectal Exam
– Biopsy
– PSA serum reading
PSA Serum Screen
• PSA is a biomarker for CaP
• Our Question: Does PSA play a role in
CaP?
-Mouse studies
PSA mouse study
• Mice don’t have PSA
• We already put WT PSA gene into mice
• We now need a mutant form of PSA gene to
put in mice so that we can compare whether
the mut mouse will develop CaP in a
different way than the WT PSA mouse
Project Goal
• Mutate the PSA gene (in a plasmid) nucleotide
sequence so that a Serine amino acid is changed to
an Alanine in the PSA protein (gene product)
• Prepare the mutated PSA gene for injection into
the mouse
SDM
EcoRI
TCT = Serine
Promoter
WT PSA Ser 189
SV40 Poly A
KpnI
NotI
EcoRI
EcoRI
Promoter
Site Directed
Mutagenesis
Mut PSA Ser 189 to Ala
SV40 Poly A
KpnI
NotI
EcoRI
GCT = Alanine
Tools
Restriction Digestion of DNA
5000 bp
6000 bp
Direction
of DNA
travel
250 bp
References
• Hricik, Todd
• http://www.webbooks.com/MoBio/Free/Ch9G.htm
• http://en.wikipedia.org/wiki/Sitedirected_mutagenesis
• http://www.ehealthmd.com/library/prostater
emoval/PR_glossary.html
Acknowledgements
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Todd Hricik
Dr. Sat Bhattacharya
Sven
Ross
Neil