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Date of Award
Restricted Thesis: Campus only access
Bachelor of Science
Dr. Valerie M. Kish
The goal of this research was to develop a p53 functional assay using p53 from the U373 human glioblastoma cell line. p53 is a tumor suppressor gene/protein that when inactivated, is a frequent precursor to malignancy. Pure, intact, mRNA was isolated from these cell lines and p53 eDNA was generated from RT-PCR p53 was then ligated into the pCR3.1 vector, transformed into E. coli cells, and the plasmid DNA was sequenced. A more useful vector was discovered, pTRE2, for it contains a tetracycline-responsive element that shows whether the gene it contains is on or off. Therefore, the p53 was taken out of the old vector using restriction enzymes and ligated into pTRE2. The plasmid was transformed into E. coli cells and cloned. Further work will include transfecting the plasmid into p53-null astrocytoma cells, treating them with tetracycline, and assessing the function of p53 in these cell lines using a Luciferase assay. These studies will provide the molecular basis for clinical testing of glioblastoma treatments targeted at p53 manipulation.
Larsen, Lorraine C., "Development of a p53 functional assay for use in the U373 human glioblastoma cell line" (2001). Honors Theses. 585.