用直接测定基因组DNA序列的方法检测β-地中海贫血基因突变

本文报道一种结合聚合酶链反应(PCR)技术直接测定基因组DNA中单考贝基因片段序列的方法,以及利用这种方法测定两例β-地贫纯合子的β珠蛋白基因序到结果。测定出基因点突变,一例为编码子17(A→T)突变纯合子,另一例为编码子69(G→A)突变纯合子。针对上述两个点突变合成寡核苷酸片段,末端标记~(82)P后为探针进行斑点杂交的结果与测序结果一致。

Direct-Genomic DNA Sequencing Technique and It's Application to Detecting β-Thalassemia Mutations

Direct sequencing of specific regions in genomic DNA became feasible with the invention of polymerase chain reaction (PCR) which permits amplification of specific regions of DNA. The genomic DNAs of 2 patients with β-thalassemia were studied by means of PCR and direct sequencing technique. Case 1 was found to be a codon 17 (A→T) mutation homozygote. Case 2 was found to be a codon 69 (G→A) mutation homozygote that was a previously undescribed mutation. The results were confirmed by dot hybridization with synthetic specific allele oligo nucleotideprobes.