Simultaneous genotyping of single nucleotide polymorphisms in the IL-1 gene complex by multiplex polymerase chain reaction-restriction fragment length polymorphism.
The interleukin-1 (IL-1) gene complex consists of the IL-1alpha, IL-1beta and IL-1 receptor antagonist genes. Single-nucleotide polymorphisms (SNP) in all three genes have been associated with human diseases. In this study, primers containing mismatches at 1-3 nucleotide positions were designed to incorporate a restriction site for endonuclease AlwNI or XcmI in the presence of allele-specific nucleotides at the polymorphic positions. Based on this technique, a simple and robust multiplex polymerase chain reaction/restriction fragment length polymorphism (multiplex PCR/RFLP) assay was developed to determine simultaneously three to four informative SNPs (IL-1beta/+3954, IL-1beta/-511 and IL-1Ra/9261 or IL-1alpha/-889, IL-1beta/-31, IL-1beta/5810 and IL-1Ra/11100 SNPs) in the IL-1 gene complex.
Tseng
Li-Hui
LH
Department of Medical Genetics, Pathology and Oncology, National Taiwan University Hospital, National Taiwan University, Taipei, Taiwan.
Chen
Pei-Jer
PJ
Lin
Ming-Tseh
MT
Singleton
Kentner
K
Martin
Emily G
EG
Yen
Ai-Hsin
AH
Martin
Paul J
PJ
Hansen
John A
JA
D000483:Alleles; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D005838:Genotype; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1 / Q000235:genetics*; D005810:Multigene Family; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D012795:Sialoglycoproteins / Q000235:genetics
D017931:DNA Primers; C507763:IL1RN protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1; D012795:Sialoglycoproteins