Baccouche Sami, Mabrouk Imed, Said Salem, Mosbah Ali, Jlidi Rachid, Gargouri Ali
Laboratory Génétique Moléculaire des Eucaryotes, Centre de Biotechnologie de Sfax, BP'K' 3038, Sfax, Tunisia.
Cancer Lett. 2003 Jan 10;189(1):91-6. doi: 10.1016/s0304-3835(02)00405-6.
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.
TP53基因第72位密码子的多态性因其在肿瘤发生和杂合性缺失（LOH）检测中的作用而得到广泛研究。通常，通过聚合酶链反应（PCR）对从血液和肿瘤组织中提取的DNA进行扩增TP53基因的外显子4，然后用AccII进行消化。在杂合性的情况下，比较血液和肿瘤DNA PCR产物的AccII图谱可以确定TP53基因座中潜在的杂合性缺失。这种方法可能会受到AccII部分消化和/或非肿瘤细胞DNA污染的阻碍。为了解决这些问题，我们开发了一种新方法，利用外显子4和外显子6之间的AccII限制性位点。外显子4-6的PCR扩增，随后进行AccII消化，使我们能够明确检测到任何杂合性缺失情况。
