摘要
To determine new polymorphisms in the antioxidant enzymes superoxide dismutase, glutathione peroxidases, catalase, and microsomal glutathione transferase 1, a search of the human expressed sequence tags (EST) database was performed (with BLAST 2.0). When any mutation, indicated by the BLAST search, gave rise to a nonconservative amino acid change we performed polymerase chain reaction (PCR) restriction analysis and/or sequence analysis of genomic DNA from human subjects in order to verify these potential polymorphisms. Of nine indicated polymorphisms from the EST analysis found in four different antioxidant enzymes, we could verify one, an amino acid substitution Pro-Leu at amino acid position 197 (P197L), in the glutathione peroxidase 1 gene. The corresponding allele frequencies were ≈70/30%. In addition, a silent mutation (1167T/C) in the catalase gene indicated by the BLAST search could also be verified. Six to nine individuals were analyzed per indicated polymorphism, so that only common polymorphisms would be found. The indicated mutations not verified by direct analysis thus cannot be excluded as allelic variation in the human population. These results show that the EST database can be used to search for polymorphisms in genes with high abundance in the human EST database. In addition to the EST analysis, PCR/single-strand conformation polymorphism (SSCP) was employed for the analysis of the microsomal glutathione transferase 1 gene. No polymorphism in the coding sequence could be detected in the gene by either method. The high degree of conservation of the microsomal glutathione transferase 1 gene indicates an important physiological function for this enzyme. Hum Mutat 13:294–300, 1999. © 1999 Wiley-Liss, Inc. .overlined { text-decoration: overline; } .struck { text-decoration:line-through; } .underlined { text-decoration:underline; } .doubleUnderlined { text-decoration:underline;border-bottom:1px solid #000; } 3.0.CO;2-5/epdf">Get PDF (312K)3.0.CO;2-5/pdf">Get PDF (312K) More content like thisFind more content: 3.0.CO;2-5&scope=allContent&start=1&resultsPerPage=20">like this articleFind more content written by: Lena Forsberg Ulf de Faire Ralf Morgenstern All Authors
摘要译文
为了确定抗氧化酶超氧化物歧化酶，谷胱甘肽过氧化物酶，过氧化氢酶和微粒体谷胱甘肽转移酶1的新多态性，对人类表达的序列标签（EST）数据库进行搜索（使用BLAST 2.0）。当由BLAST搜索指示的任何突变时，为了验证这些潜在的多态性，我们对来自人类受试者的基因组DNA进行聚合酶链式反应（PCR）限制性分析和/或序列分析。在来自在四种不同抗氧化酶中发现的EST分析的9个多态性中，我们可以证实一个氨基酸取代Pro-Leu位于第197位氨基酸（P197L），在谷胱甘肽过氧化物酶1基因中。相应的等位基因频率是≈70/ 30％。此外，BLAST搜索所示的过氧化氢酶基因中的沉默突变（1167T / C）也可以被验证。根据指示的多态性分析六至九个个体，所以只能找到常见的多态性。未被直接分析证实的所示突变因此不能被排除为人类中的等位基因变异。这些结果表明，EST数据库可用于在人类EST数据库中搜索高丰度基因的多态性。除EST分析外，采用PCR /单链构象多态性（SSCP）分析微粒体谷胱甘肽转移酶1基因。用这两种方法都不能在基因中检测到编码序列的多态性。微粒体谷胱甘肽转移酶1基因的高度保守性表明该酶的重要生理功能。 Hum Mutat 13：294-300,1999。©1999 Wiley-Liss，Inc.。[overlined {text-decoration：overline; } .struck {text-decoration：line-through; } .underlined {text-decoration：underline; } .doubleUnderlined {text-decoration：underline; border-bottom：1px solid＃000;][} 3.0.CO; 2-5 / epdf“获取PDF（312K）3.0.CO; 2-5 / pdf”获取PDF（312K）更多类似产品的内容查看更多内容：3.0.CO;]2-5＆scope \x3d allContent＆start \x3d 1＆resultsPerPage \x3d 20“像这篇文章查找更多内容撰写者：Lena Forsberg Ulf de Faire Ralf Morgenstern所有着者
Lena Forsberg[1,*]; Ulf de Faire[2] and Ralf Morgenstern[1];. Low yield of polymorphisms from EST blast searching: Analysis of genes related to oxidative stress and verification of the P197L polymorphism in GPX1[J]. Human Mutation, 1999,13(4): 294-300