Biochemical and Biophysical Research Communications
Genetic polymorphism of the human cytochrome CYP2A13 in a French population: implication in lung cancer susceptibility
Section snippets
Materials and methods
Genomic DNA samples. A group of 102 unrelated French Caucasian subjects has been involved in the study after Ethical Committee approval and informed consents have been obtained. Among the 102 subjects, 76 were healthy volunteers and 26 were patients with various bowel diseases. Nucleic acid extraction from peripheral blood leucocytes was performed using the Nucleon BACC3 kit (Amersham–Pharmacia Biotech, Saclay, France), according to manufacturer's instructions.
Additionally, genotyping of the
Sequence analysis of CYP2A13 in a French population
To investigate the extent of CYP2A13 gene polymorphism in a large French Caucasian population, a PCR-SSCP strategy was applied to genomic DNAs of 102 unrelated volunteers. One DNA sample homozygote for a wild-type allele of CYP2A13, as confirmed by sequencing, was used as a reference for each SSCP analysis. Single strands were well separated and easily discriminated for each of the CYP2A13 regions studied, as illustrated in Fig. 1 for exons 2, 3, 4, 6, and 9.
For 80 samples, electrophoretic
Discussion
In order to carry out a screening for sequence variations in the 5′-flanking and coding regions of the CYP2A13 gene, a PCR-SSCP strategy was applied to 102 unrelated French individuals of Caucasian origin [17]. In the past decade, this method has been successfully used for the detection of mutations in several human cytochrome P450 genes [18]. After optimization of several parameters affecting the single-strand separation, such as temperature for migration or acrylamide concentration, we
Acknowledgements
This study was supported by the Centre Hospitalier Régional et Universitaire de Lille, France, the Fondation pour la Recherche Médicale, France, and by the Génopole de Lille, Région Nord-Pas de Calais (FEDER), France.
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