A more accurate detection of codon 72 polymorphism and LOH of the TP53 gene.
The polymorphism at codon 72 of the TP53 gene has been extensively studied for its involvement in cancerogenesis and loss of heterozygosity (LOH) detection. Usually, the exon 4 of the TP53 gene is amplified by polymerase chain reaction (PCR) on DNA extracted from blood and tumor tissues, then digested by AccII. In the case of heterozygosity, the comparison of AccII profile from blood and tumor DNA PCR products allowed the identification of a potential LOH in the TP53 locus. This method can be hindered by a partial AccII digestion and/or DNA contamination of non-tumor cells. To circumvent these problems, we have developed a new approach by using the AccII restriction site between exon 4 and exon 6. The PCR amplification of exon 4-6, followed by AccII digestion allowed us to detect without ambiguity any LOH case.
Baccouche
Sami
S
Laboratory Génétique Moléculaire des Eucaryotes, Centre de Biotechnologie de Sfax, BP'K' 3038, Sfax, Tunisia.
Mabrouk
Imed
I
Said
Salem
S
Mosbah
Ali
A
Jlidi
Rachid
R
Gargouri
Ali
A
D004252:DNA Mutational Analysis / Q000379:methods*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics; D016158:Genes, p53; D006801:Humans; D019656:Loss of Heterozygosity; D009369:Neoplasms / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012680:Sensitivity and Specificity
C039566:CGCG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific