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目的:鉴定1例葡萄糖-6-磷酸脱氢酶缺乏症患者的基因突变。方法:用聚合酶链反应、限制性内切酶筛查葡萄糖-6-磷酸酶基因1388G→A、1376G→T、1360C→T、1024C→T、592C→T、517T→C、493A→G,487G→A、392G→T、95A→G突变,用单链构象多态性筛查葡萄糖-6-磷酸脱氢酶基因的所有外显子,用核苷酸序列测定确定基因突变。结果:该患者未存在1388G→A、1376G→T、1360C→T、1024C→T、592C→T、517T→C、493A→G、487G→A、392G→T、95A→G突变,但在外显子8发现了一种新的G6PD基因突变———835A→G突变,此突变导致第279位的苏氨酸被丙氨酸取代,将其命名为G6PD-海口,其酶活性约是正常的10%,比835A→T突变型的活性低,后者的酶活性约是正常的40%;分析人G6PD的三维结构模型表明,第279位苏氨酸残基的羟基对于维持G6PD亚基的相互作用具有非常重要的作用。结论:835A→G突变是一种新的G6PD基因突变型,G6PD的第279位苏氨酸残基的羟基是维持G6PD亚基相互作用及酶活性的必需基团。
Objective: To identify a gene mutation in a patient with glucose-6-phosphate dehydrogenase deficiency. METHODS: The glucose-6-phosphatase gene 1388G → A, 1376G → T, 1360C → T, 1024C → T, 592C → T, 517T → C and 493A → G were screened by polymerase chain reaction and restriction endonuclease. 487G → A, 392G → T, 95A → G mutations. All exons of glucose-6-phosphate dehydrogenase gene were screened by single-strand conformation polymorphism, and the gene mutation was determined by nucleotide sequence determination. Results: There were no 1388G → A, 1376G → T, 1360C → T, 1024C → T, 592C → T, 517T → C, 493A → G, 487G → A, 392G → T, 95A → G mutations in this patient, Son 8 found a new G6PD gene mutation --- 835A → G mutation, this mutation led to the 279th threonine replaced by alanine, named G6PD-Haikou, the enzyme activity is about normal 10%, which is lower than the activity of 835A → T mutant, and the activity of the latter is about 40% of normal. Analysis of the three-dimensional structure model of human G6PD shows that the hydroxyl group of the threonine residue at position 279 is not able to maintain the G6PD subunit Interaction has a very important role. CONCLUSION: The 835A → G mutation is a new mutant of G6PD gene. The hydroxyl group of threonine residue at position 279 of G6PD is an essential group to maintain the G6PD subunit interaction and enzyme activity.