Complete sequencing of a genetic polymorphism in NAT2 in the Korean population.
Complete sequencing of a genetic polymorphism in NAT2 in the Korean population.
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DOI：
10.1093/clinchem/48.5.775
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发表时间：
2002-05
影响因子：
9.3
通讯作者：
Soo-Youn Lee;Kyung‐A Lee;C. Ki;O. J. Kwon;Ho Joong Kim;M. Chung;G. Suh;Jong-Won Kim
中科院分区：
文献类型：
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作者：
Soo-Youn Lee;Kyung‐A Lee;C. Ki;O. J. Kwon;Ho Joong Kim;M. Chung;G. Suh;Jong-Won Kim
N -乙酰转移酶2（NAT 2）代谢芳胺和肼。NAT 2的底物包括许多治疗药物，如异烟肼（INH），以及化学品和致癌物（1）（2）（3）。因此，N-乙酰化活性与药物作用或毒性以及对各种癌症的易感性相关。NAT 2使芳胺N-乙酰化的能力受NAT 2基因中的遗传多态性的影响。乙酰化率和NAT 2基因型分布在不同群体间差异较大。NAT 2的遗传多态性在韩国人群中尚未得到广泛研究。以前的报告仅基于表型或限制性片段长度多态性分析，导致可能对基因型进行错误分类。因此，我们决定通过完全测序来调查韩国人群中NAT 2等位基因变异和基因型分布。我们还评估了基因型和表型之间的关系，以了解N-乙酰化药物遗传学。对1000名访问三星医疗中心健康促进中心的韩国人进行了匿名研究。另外23名健康志愿者和18名肺结核患者参加了这项研究。从外周血白细胞中提取DNA。我们扩增了一个1211 bp的片段，其中包括NAT 2基因的870 bp蛋白编码区，并在ABI Prism 377 DNA测序仪（Perkin-Elmer）上进行了全测序分析（4）。然后，我们检查核苷酸替换等位基因特异性PCR（AS-PCR）和限制性内切酶消化的组合使用。在单独的PCR中使用野生型和突变等位基因的特异性引物（5）来检测C282 T、T341 C和G590 A置换。通过分别用Nci I、Kpn I、Taq I、Dde I和Bam HI消化携带野生型或突变体等位基因的1211-bp PCR片段或AS-PCR产物来探索位置190、481、590、803和857处的核苷酸。对于表型，健康志愿者和患者提供了知情的...
N -Acetyltransferase 2 (NAT2) metabolizes arylamines and hydrazines. The substrates of NAT2 include many therapeutic drugs, such as isoniazid (INH), as well as chemicals and carcinogens (1)(2)(3). For that reason, N-acetylation activity is associated with drug effects or toxicities and susceptibility to various cancers. The ability of NAT2 to N-acetylate arylamines is subject to a genetic polymorphism in the NAT2 gene. The acetylation rate and NAT2 genotype distribution are quite different among various populations. The genetic polymorphism in NAT2 has not been studied extensively in the Korean population. Previous reports were based only on phenotyping or restriction fragment length polymorphism analysis, leading to possible misclassification of genotypes. We therefore decided to investigate NAT2 allelic variability and genotype distributions in the Korean population by complete sequencing. We also evaluated the relationship between genotype and phenotype to understand N-acetylation pharmacogenetics. One thousand Korean individuals who visited the health promotion center at Samsung Medical Center were anonymously studied. An additional 23 healthy volunteers and 18 patients with pulmonary tuberculosis participated in this study. DNA was extracted from peripheral blood leukocytes. We amplified a 1211-bp fragment that included an 870-bp protein-coding region of the NAT2 gene and performed full sequencing analysis (4) on an ABI Prism 377 DNA Sequencer (Perkin-Elmer). We then checked nucleotide substitutions by combined use of allele-specific-PCR (AS-PCR) and restriction enzyme digestion. Specific primers (5) for the wild-type and mutant alleles were used in separate PCRs to detect C282T, T341C, and G590A substitutions. The nucleotides at positions 190, 481, 590, 803, and 857 were explored by digesting the 1211-bp PCR fragment or AS-PCR product carrying the wild-type or mutant allele with Nci I, Kpn I, Taq I, Dde I, and Bam HI, respectively. For phenotyping, the healthy volunteers and patients gave informed …