Genetic typing of the human cytochrome P450 2A6 gene and related methods

Abstract

Disclosed are novel polymorphisms in the human cytochrome P450 2A6 gene and the use of those polymorphisms as predictive sequences for altered metabolism or occurrence of disease.

1 - 35 . (canceled)  
     
     
         36 . A method of detecting the presence of a G polymorphism at promoter position −39 of a CYP2A6 gene in an individual, the method comprising: 
 (a) obtaining a nucleic acid sample from the individual; and    (b) amplifying a region of the CYP2A6 gene using a forward primer that hybridizes to the gene upstream of promoter position −39 and a reverse primer that hybridizes to intron 1 of the gene to generate an amplification product; and    (c) assaying the amplification product to determine whether the individual has a G polymorphism at promoter position −39 of the CYP2A6 gene.    
     
     
         37 . The method of  claim 36 , wherein assaying comprises determining whether the individual is homozygous or heterozygous for the G polymorphism.  
     
     
         38 . The method of  claim 36 , wherein the reverse primer has a nucleotide sequence comprising contiguous nucleotides from intron 1.  
     
     
         39 . The method of  claim 36 , wherein the forward primer comprises SEQ ID NO:1 and the reverse primer comprises SEQ ID NO:2.  
     
     
         40 . A method of detecting the presence or absence of a G polymorphism at promoter position −39 of a CYP2A6 gene in an individual, the method comprising: 
 (a) obtaining a nucleic acid sample from the individual; and    (b) mixing the nucleic acid sample with an allele-specific primer, specific for a G polymorphism or a T at promoter position −39 of the CYP2A6 gene, such that upon hybridization a polymerase-mediated extension product forms if an allelic form complementary to the allele-specific primer is present; and    (c) analyzing the extension product to identify the presence or absence of the G polymorphism at promoter position −39.    
     
     
         41 . The method of  claim 40 , wherein the method further comprises contacting the nucleic acid sample with a second primer which hybridizes to a distal site such that an allele-specific amplification product is formed.  
     
     
         42 . The method of  claim 40 , wherein analyzing comprises determining whether the individual is homozygous or heterozygous for the G polymorphism.  
     
     
         43 . A method of detecting the presence of a G polymorphism at promoter position −39 of a CYP2A6 gene in an individual, the method comprising: 
 (a) obtaining a nucleic acid sample from the individual; and    (b) generating an amplification product from the nucleic acid sample specific for a region of the CYP2A6 gene spanning promoter position −39, wherein the amplified region lacks exonic sequences of the CYP2A6 gene; and    (c) analyzing the amplification products to identify the presence or absence of the G polymorphism    
     
     
         44 . The method of  claim 43 , wherein analyzing comprises determining whether the individual is homozygous or heterozygous for the G polymorphism.  
     
     
         45 . The method of  claim 43 , wherein an allele-specific primer for promoter position −39 of the CYP2A6 gene is used to amplify the region.