葡萄糖-6-磷酸脱氢酶基因突变与NlaⅢ多态性位点连锁关系研究

Linkage analysis of the G6PD gene mutations and its NlaⅢ polymorphic site

目的 研究葡萄糖 6 磷酸脱氢酶 (G6PD)基因内的特定多态性位点与G6PD缺陷基因突变的连锁关系 ,进一步揭示G6PD缺陷症分子病理学本质及G6PD基因的多态性位点在人类学和人类迁移研究中的意义。方法 应用变性梯度凝胶电泳和DNA点杂交技术检测 5 4例G6PD活性正常男性对照者及 6 6例男性G6PD缺陷者G6PD基因中距离 12外显子上游 13bp处的NlaⅢ多态性位点。结果 在 6 6例G6PD缺陷者中 1376 (G→T)突变 32例、1388(G→A)突变 2 1例、95 (A→G)突变 13例。G6PD活性正常对照者中 ,距离G6PD基因 12外显子上游 13bp处的碱基是T(- 13bpexon12T ,无NlaⅢ内切酶位点 )者共 11例 ,约占 2 0 .4% ;而该位点碱基是C(- 13bpexon12C ,有NlaⅢ内切酶位点 )者共 43例 ,占 79.6 %。 6 6例G6PD缺陷者均与 - 13bpexon12C连锁遗传。结论 G6PD基因突变型 1376 (G→T)、1388(G→A)、95 (A→G)与 - 13bpexon12C连锁遗传是中国人G6PD缺陷症的特征之一。这一特征对于研究G6PD缺陷症复杂的临床表现有一定意义 ,对于人类学研究有参考作用。

Objective To investigate the molecular and anthropologic features of glucose 6 phosphate dehydrogenase (G6PD) deficiency and the linkage between the Nla Ⅲ polymorphic site within G6PD gene and three common Chinese G6PD mutations. Methods By using denatured gradient gel electrophoresis(DGGE) and allele specific oligonucleotide(ASO) probe dot blot hybridization, Nla Ⅲ polymorphic site at -13 bp upstream exon 12 within G6PD gene were screened in 54 males with normal G6PD activity and 66 G6PD deficient males.Results Thirty two cases with cDNA1376(G→T),21 with cDNA1388(G→A) and 13 with cDNA95(A→G) were detected in the 66 G6PD deficient males. Among 54 normal controls, T at -13 bp upstream exon 12 within G6PD gene were detected in 11 cases (20.4%, without Nla Ⅲ polymorphic sites) while C in 43 cases (79.6%,with Nla Ⅲ polymorphic sites). All 66 G6PD deficient males were linked with C at -13 bp upstream exon 12 (with Nla Ⅲ polymorphic sites). Conclusion The G6PD mutations cDNA1376 (G→T), cDNA1388(G→A) and cDNA95(A→G) were linked with Nla Ⅲ polymorphism. This feature will play a role in studying the complicated manifestations and anthropology of G6PD deficiency.