title: Simple genotype analysis of the Asp299Gly polymorphism of the Toll-like receptor-4 gene that is associated with lipopolysaccharide hyporesponsiveness.
abstract: A nonsynonymous single nucleotide polymorphism (Asp299Gly) in the Toll-like receptor-4 (TLR-4) gene affects the responsiveness to lipopolysaccharide in humans. To analyze this important polymorphism more efficiently, we developed a simple polymerase chain reaction (PCR) restriction length fragment polymorphism (RFLP) assay and examined the Asp299Gly allele frequency in a Japanese population. The PCR primer was designed with 1- or 2-bp mismatches, creating the recognition sequence for restriction enzyme BsaBI or BstXI, allowing RFLP analysis of the digested products. Genotyping was carried out with this assay for 275 DNA specimens from 107 healthy volunteers and 168 patients with various diseases, including ulcerative colitis (n = 86). The Asp299Gly allele of the TLR-4 gene was not detected in any of the specimens, suggesting that it is very rare in Japanese.
authors.affiliation: Division of Clinical Laboratory, Yamaguchi University Hospital, Yamaguchi University School of Medicine, Yamaguchi, Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Okayama
authors.forename: Kozue
authors.initials: K
authors.lastname: Fujimura
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Suehiro
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Hamanaka
authors.forename: Motoki
authors.initials: M
authors.lastname: Fujiwara
authors.forename: Tomoyo
authors.initials: T
authors.lastname: Matsubara
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Maekawa
authors.forename: Shoichi
authors.initials: S
authors.lastname: Hazama
authors.forename: Masaaki
authors.initials: M
authors.lastname: Oka
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Nohara
authors.forename: Kozo
authors.initials: K
authors.lastname: Kayano
authors.forename: Kiwamu
authors.initials: K
authors.lastname: Okita
authors.forename: Yuji
authors.initials: Y
authors.lastname: Hinoda
mesh_terms: D003093:Colitis, Ulcerative / Q000097:blood / Q000235:genetics*; D029721:Drosophila Proteins; D005838:Genotype; D006801:Humans; D008070:Lipopolysaccharides / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000097:blood / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D011956:Receptors, Cell Surface / Q000097:blood / Q000235:genetics*; D051197:Toll-Like Receptor 4; D051193:Toll-Like Receptors
chemical_list: D029721:Drosophila Proteins; D008070:Lipopolysaccharides; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4; D051193:Toll-Like Receptors