摘要
目的 了解中国人苯丙酮尿症 ( phenylketonuria,PKU)患者的苯丙氨酸羟化酶( phenylalanine hydroxylase,PAH)基因第 12外显子点突变种类和频率。方法 应用单链构象多态性( single strand conformation polymorphism,SSCP)、变性梯度凝胶电泳 ( denaturing gradient gelelectrophoresis,DGGE)、DNA测序分析了 12 7例 PKU患者的 PAH基因第 12外显子点突变种类及频率。结果 DNA测序分析显示 10例患者存在 R4 13P、S4 11X、R4 0 8W、R4 0 8Q 4种杂合突变 ,其突变频率分别为 2 .76 %、0 .39%、0 .39%、0 .39% ,S4 11X突变为中国人中首次报道。 SSCP分析仅发现 2例 R4 13P杂合突变 ,DGGE分析显示 10例出现 3种类型的异常电泳带型。R4 13P突变在南北方人之间、在经典型 PKU和高苯丙氨酸血症之间的分布差异无显著性。结论 DGGE对 PAH基因第 12外显子点突变检出率明显高于 SSCP。 DGGE结合 DNA测序是明确 PAH基因第 12外显子点突变种类和频率较好的方法。 R4
Objective To obtain the mutation spectrum of exon 12 of the phenylalanine hydroxylase( PAH )gene. Methods The samples from 127 patients with phenylketonuria(PKU) were studied by polymerase chain reaction(PCR)-single strand conformation polymorphism(SSCP),PCR-denaturing gradient gel electrophoresis(DGGE) and direct DNA sequencing. Results Four heterozygous mutations including R413P,S411X,R408W,R408Q in 10 of 127 patients were identified by DNA sequencing. The frequencies of R413P,S411X,R408W,R408Q mutation were 2.76%, 0.39%, 0.39% and 0.39%, respectively. The S411X mutation in Chinese was first reported. Only 2 cases with R413P mutation were detected by SSCP. DGGE analysis showed that 10 cases displayed 3 kinds of abnormal electrophoretic bands. There were no obvious differences in the frequency of R413P mutation between northern and southern Chinese with PKU, and between classical PKU and hyperphenylalaninemia. Conclusion DGGE is more sensitive than SSCP in screening the mutations of exon 12 of the PAH gene. DGGE combined with DNA sequencing can be used to define all the mutations of exon 12 of the PAH gene. These results indicate the similarity in the frequency of R413P mutation between northern and southern patients.
出处
《中华医学遗传学杂志》
CAS
CSCD
2004年第3期261-263,共3页
Chinese Journal of Medical Genetics
关键词
苯丙氨酸羟化酶基因
基因突变
单链构象多态性
变性梯度凝胶电泳
苯丙酮尿症
phenylalanine hydroxylase gene
gene mutation
single strand conformation polymorphism
denaturing gradient gel electrophoresis
