Duplex Detection of TP53 Arg72Pro and 16 bp Del/Ins Polymorphisms by a Simple Optimized PCR-RFLP Method.
BACKGROUND
The tumor suppressor gene (TP53) encodes p53, the central protein in the apoptotic pathway which has been shown to be of crucial importance in the development of cancers in addition to a variety of neurodegenerative disorders. Two most commonly studied polymorphisms that were shown to affect the biochemical functions of p53 protein are the exon 4 Arg72pro and Intron 3 16 bp Del/Ins polymorphisms.


OBJECTIVE
The aim of the present work is to develop a new optimized method for the simultaneous detection of the two important polymorphisms in the TP53 gene in a single reaction.


METHODS
The proposed method is based on amplification of a single PCR amplicon and the use of a unique restriction enzyme with restriction sites that facilitate simultaneous detection.


RESULTS
The proposed method offers fast, economical, and simple simultaneous detection. Validation by methods commonly used in the literature showed perferct concordance in genotyping results.


CONCLUSIONS
The proposed method can serve as an invaluable tool for the investigation of TP53 Arg72Pro-16 bp Del/Ins haplotype, and the combined effects of the two polymorphisms offering extreme ease and simplicity over the currently used methods which are based on two separate detections.
Department of Analytical Chemistry, Faculty of Pharmacy, University of Aleppo, Aleppo, Syria.
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B
Lajin
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Alachkar
Sakur A
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Alhaj