Hb D Punjab基因的鉴定——DNA 扩增在异常血红蛋白研究中的应用

IDENTIFICATION OF Hb D-PUNJAB: AN APPLICATION OF DNA AMPLIFICATION ON STUDY OF ABNORMAL HEMOGLOBINS

本文应用作者设计的寡核苷酸引物和探针,通过聚合酶链反应(PCR)扩增β珠蛋白DNA序列,并通过限制酶EcoRI酶谱分析或寡核苷酸杂交,快速鉴定Hb D Punjab基因。应用这种技术先后对汉、藏、哈萨克等3个民族4个家系的Hb D Punjab基因进行了鉴定。

Hemoglobin D-Punjab is a common Hb variant in China. This paper describes a new way——Eco RI mapping of the amplified β-globin DNA, for identification of Hb D-Punjab gene. The primers for PCR were designed and synthesized to enzymatically amplify an 144bp fragment which contained an Eco R1 recognition site. So the D-Punjab gene could be easily detected by Eco RI digestion of the amplified sequences on agarose gel electrophoresis owing to a single base change on codon 121. Four Hb DPunjab families from Han, Tibet and Kasak nationalities were analysed by this simple method. The results were also confirmed by oligonucleotide hybridization technique.