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Mouse Models of Prostate Cancer
Name: Vicki Javier
Mentor: Todd Hricik
Institution: MSKCC
• 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
Prostate Cancer
• PSA is a biomarker for CaP
• Our Question: Does PSA play a role in CaP?-Mouse studies
PSA Serum Screen
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
• 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
Project Goal
Promoter WT PSA Ser 189 SV40 Poly A
NotIKpnI
EcoRI
EcoRI
Promoter Mut PSA Ser 189 to Ala SV40 Poly A
NotIKpnI
EcoRI
EcoRI
SDM
Site Directed Mutagenesis
TCT = Serine
GCT = Alanine
Tools
Restriction Digestion of DNA
Direction of DNA travel
6000 bp5000 bp
250 bp
References
• Hricik, Todd
• http://www.web-books.com/MoBio/Free/Ch9G.htm
• http://en.wikipedia.org/wiki/Site-directed_mutagenesis
• http://www.ehealthmd.com/library/prostateremoval/PR_glossary.html
Acknowledgements
• Todd Hricik
• Dr. Sat Bhattacharya
• Sven
• Ross
• Neil