title: Application of CRS-PCR-RFLP to identify CYP1A1 gene polymorphism.
abstract: BACKGROUND
  Cytochrome P4501A1 (CYP1A1) is a member of the cytochrome P450 gene family and plays an important role in the metabolism of exogenous and endogenous material. In recent research, it has been shown that its genetic polymorphisms are associated with many diseases. But the isoschizomers such as the BsrDI enzyme required for the detection of this polymorphism are expensive.
  
  
  METHODS
  The study used an improved PCR-RFLP method with mismatched base for detection of the single-nucleotide polymorphism rs1048943. A new restriction enzyme cutting site was created by created restriction site PCR (CRS-PCR), and the restriction enzyme StyI for RFLP was cheaper than other enzymes. A total of 320 samples from Han Chinese were tested to evaluate this novel method. The PCR results were confirmed by DNA sequencing.
  
  
  RESULTS
  After detecting 320 Chinese Han individuals, the genotype frequencies were 63.74% for AA, 31.54% for AG, and 4.72% for GG. The allelic frequencies were 75.48% for A and 24.52% for G. The x2test showed the genotype and allele frequencies of CYP1A1 do not deviate from Hardy-Weinberg equilibrium, and the sequences of amplified products were consistent with the one published in GenBank with the exception of mismatched base.
  
  
  CONCLUSIONS
  Based on PCR with mismatched primers we designed, the CYP1A1 polymorphism could be identified effectively with low cost.
authors.affiliation: Department of Occupational Health and Occupational Disease, College of Public Health, Zhengzhou University, Zhengzhou, China.
authors.forename: Mingcui
authors.initials: M
authors.lastname: Ding
authors.affiliation: Department of Environmental Health, College of Public Health, Zhengzhou University, Zhengzhou, China.
authors.forename: Xiaoran
authors.identifier: http://orcid.org/0000-0001-8533-7750
authors.initials: X
authors.lastname: Duan
authors.affiliation: Department of Occupational Health and Occupational Disease, College of Public Health, Zhengzhou University, Zhengzhou, China.
authors.forename: Xiaolei
authors.initials: X
authors.lastname: Feng
authors.affiliation: Department of Occupational Health and Occupational Disease, College of Public Health, Zhengzhou University, Zhengzhou, China.
authors.forename: Pengpeng
authors.initials: P
authors.lastname: Wang
authors.affiliation: Department of Occupational Health and Occupational Disease, College of Public Health, Zhengzhou University, Zhengzhou, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
mesh_terms: D044466:Asian People / Q000235:genetics; D002681:China; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics*; D004247:DNA / Q000032:analysis / Q000235:genetics; D005787:Gene Frequency / Q000235:genetics; D006801:Humans; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*
chemical_list: D004247:DNA; D019363:Cytochrome P-450 CYP1A1
keywords: Cytochrome P4501A1; PCR-RFLP; Single-nucleotide polymorphism; created restriction site; rs1048943