title: Investigation of the -1377 polymorphism on the Apo-1/Fas promoter in systemic lupus erythematosus patients using allele-specific amplification.
abstract: Apoptosis mediated by the Apo-1/Fas and Fas ligand pathways has been implicated in many disorders, including autoimmunity and tumorigenesis. The recent identification of two polymorphisms on the 5' flanking region of the human Apo-1/Fas gene has provided useful markers for investigation of the genetic contribution of the Apo-1/Fas gene in these diseases. The Mval polymorphism at the -670 nucleotide position is frequent in the normal population, with 51% heterozygosity. The other polymorphism, a result of single nucleotide G-->A substitution at the -1377 position, does not create or delete any restriction enzyme digestion sites. In this paper, we describe a simple and rapid method for detecting the -1377 polymorphism by using allele-specific amplification (ASA). Using the ASA method, the -1377 polymorphism in a normal Caucasian population was characterised. Frequencies of 0.13 and 0.87 for allele A and G, respectively, were observed and the homozygosity of the mutant allele (A) was found in only 2% of the population. We subsequently examined the -1377 polymorphism in sporadic systemic lupus erythematosus (SLE) patients (n = 86). The results showed that both genotype distribution and allele frequencies in SLE patients were similar to that in controls, suggesting that the -1377 promoter polymorphism is unlikely to be associated with SLE susceptibility. The description of this rapid detection method and characterisation of the -1377 polymorphism are useful means for future genetic studies in diseases in which the Fas-mediated apoptosis may be involved.
authors.affiliation: Department of Rheumatology, Westmead Hospital, University of Sydney, Australia.
authors.forename: Q R
authors.initials: QR
authors.lastname: Huang
authors.forename: N
authors.initials: N
authors.lastname: Manolios
mesh_terms: D000328:Adult; D000368:Aged; D000483:Alleles; D016632:Apolipoprotein A-I / Q000235:genetics*; D004247:DNA / Q000032:analysis; D004252:DNA Mutational Analysis; D017931:DNA Primers / Q000737:chemistry; D053222:Fas Ligand Protein; D005260:Female; D005784:Gene Amplification; D005838:Genotype; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000235:genetics*; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics*; D008875:Middle Aged; D008969:Molecular Sequence Data; D010375:Pedigree; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic
chemical_list: D016632:Apolipoprotein A-I; D017931:DNA Primers; C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D004247:DNA