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The mutations in TP53 gene from the glioma patients that trigger the aberrant pre-mRNA splicing

TP53 gene encodes the “Tumour Protein” p53 which plays a central role in cell cycle regulation. This protein is widely considered as the “Guardian of the Genome” for maintaining genetic stability. A medical interest to this protein is driven by the fact that about 50% of all human cancers contain mutations in the TP53 gene. We are screening for novel TP53 mutations in the cell line originated from the glioma patients. We have identified the single nucleotide substitution that creates the stop codon which supposed to truncate the protein to one third of its full-length. However, a “full-length” p53 is detected by western blotting in extracts from this cell line. The phenomenon is appeared to be a result of the aberrant pre-mRNA splicing which skips the stop codon.

The aims of this project are to (i) investigate the molecular mechanism of pre-mRNA splicing of the mutant TP53 gene which results in skipping the stop codon and producing a nearly full-length p53 protein; (ii) address the functional roles of the truncated and the single amino acid deletion mutants of p53 in cell cycle regulation and cancerogenesis; and (iii) continue the screening for the mutations in TP53 gene from the glioma patients that could lead to the aberrant pre-mRNA splicing. Methods: DNA cloning, tissue culture, mini-gene pre-mRNA splicing, flow cytometry, fluorescence microscopy, real-time PCR.

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