Kuwata S, Yanagisawa M, Saeki H, Nakagawa H, Etoh T, Tokunaga K, Juji T, Shibata Y
Department of Transfusion Medicine and Immunohematology, Faculty of Medicine, University of Tokyo, Japan.
J Allergy Clin Immunol. 1995 Dec;96(6 Pt 2):1051-60. doi: 10.1016/s0091-6749(95)70190-7.
We examined polymorphisms of transporter associated with antigen processing (TAP) genes in 37 Japanese patients with atopic dermatitis and 52 control subjects. We have evaluated, again, the specificity polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method since our previous report and have also developed a mismatch PCR-RFLP method for discriminating two dimorphic sites of TAP1 gene and four dimorphic sites of TAP2 gene. Amplified products from genomic DNA were digested with restriction endonucleases: Sau3A1 for TAP1 codon 333 (Ile-Val), AccI for TAP1 codon 637 (Asp-Gly), and BfaI for TAP2 codon 687 (Stop-Gln). The other sites of TAP2 gene were analyzed by mismatch PCR-RFLP: AccII for TAP2 codon 379 (Val-Ile), RsaI for codon 565 (Ala-Thr), and MspI for codon 665 (Thr-Ala). We observed three TAP1 alleles and six TAP2 alleles. Infrequent allele TAP1 C was observed in one patient and one control subject. We did not identify any differences in TAP allele frequencies between those patients with atopic dermatitis and Japanese control subjects. Analysis of TAP gene polymorphisms will provide better understanding of susceptibility loci in HLA class II-associated disease because TAP genes are located between HLA-DQB1 and HLA-DPB1 loci.
我们检测了37例日本特应性皮炎患者和52例对照者中与抗原加工相关的转运体(TAP)基因的多态性。自我们之前的报告以来,我们再次评估了特异性聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法,并且还开发了一种错配PCR-RFLP方法,用于区分TAP1基因的两个双态位点和TAP2基因的四个双态位点。用限制性内切酶消化基因组DNA的扩增产物:用Sau3A1酶切TAP1密码子333(异亮氨酸-缬氨酸),用AccI酶切TAP1密码子637(天冬氨酸-甘氨酸),用BfaI酶切TAP2密码子687(终止密码子-谷氨酰胺)。TAP2基因的其他位点通过错配PCR-RFLP分析:用AccII酶切TAP2密码子379(缬氨酸-异亮氨酸),用RsaI酶切密码子565(丙氨酸-苏氨酸),用MspI酶切密码子665(苏氨酸-丙氨酸)。我们观察到3个TAP1等位基因和6个TAP2等位基因。在1例患者和1例对照者中观察到罕见等位基因TAP1 C。我们未发现特应性皮炎患者与日本对照者之间TAP等位基因频率存在任何差异。由于TAP基因位于HLA-DQB1和HLA-DPB1位点之间,因此对TAP基因多态性的分析将有助于更好地理解HLA II类相关疾病的易感基因座。