Can I Clone the Normal GM-CSF Gene Out of My Dog's Tumor?

Overview

Objectives/Goals Last year I found multiple DNA mutations in my dogs tumor p53 growth control gene. This year I looked for mutations in the critical GM-CSF immune cancer fighting gene in my dogs tumor RNA. If my dogs tumor GM-CSF gene was mutated, perhaps her immune system was weakened and could not fight her cancer. If her tumor GM-CSF gene was normal, maybe it was expressed at low levels. Methods/Materials A)Genomic Dog tumor DNA/RNA isolation. B)The Dog GM-CSF gene was amplified using polymerase chain reaction (PCR) and ligated into a sequencing plasmid. C)The Dog GM-CSF PCR amplified gene was sequenced. D)The sequence data was analyzed using Sequencher software and electronically compared to the normal dog GM-CSF gene sequence. Results I was able to use RT-PCR to amplify the Dog tumor GM-CSF gene and clone it into a sequencing plasmid.The sequence data for my cloned Dog GMCSF gene is clear and strong and shows no mutations. Conclusions/Discussion Last year I found multiple DNA mutations in my dogs tumor p53 growth control gene which explained her tumor growth. This year I find no mutations in her tumor immune cancer fighting GM-CSF gene. Her immune system must have been fighting her cancer since GM-CSF was expressed, but maybe not expressed high enough. Perhaps I can put the normal GM-CSF in a high expression plasmid and use it as a cancer fighting vaccine for other Dogs Cancer in the future.

Competition history

• CSEF 2007 Biochemistry/ Molecular Biology · Entry J0418

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