Analysis of the non-deletion α-thalassemia mutations by PCR temperature gradient gel electrophoresis
10.3760/j.issn:1003-9406.2001.01.015
- VernacularTitle:应用温度梯度凝胶电泳分析非缺失型α-地中海贫血突变
- Author:
Yongzhong ZHAO
1
;
Xiangmin XU
;
Yang YANG
Author Information
1. the First Military Medical University
- From:
Chinese Journal of Medical Genetics
2001;18(1):51-55
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop a PCR-temperature gradient gel electrophoresis(TGGE) method applied to screening for the point mutations causing the non-deletion α-thalassemia.Methods The entire α2-globin gene fragment (1085 bp) was selectively amplified from human genomic DNA with different genotypes of α-thalassemia and a 543 bp fragment spanning the exon2 and exon3 of the α2-globin gene was amplified with a pair of nested primers. The condition of perpendicular and parallel TGGE for the two different fragments in length was optimized and the candidate mutant was confirmed by DNA sequencing. In pilot study, 15 samples with suspected non-deletion α-thalassemia were screened for point mutations in α-globin gene. Results Hb Constant Spring (Hb CS) and Hb Quong Sze (Hb QS), two most commonly types of the non-deletion α-thalassemia distributed in Chinese, and a rare type of the Hb Westmead mutation, α2 CD122 CAC→CAG(His→Gln) could be detected by PCR-TGGE. Among those 15 samples screened for α-thalassemia mutation were 10 samples with Hb CS mutation, 2 samples with Hb QS mutation, 2 samples with Hb Westmead mutation, and a previously unreported one, α2 CD31 AGG→AAG(Arg→Lys), confirmed by DNA sequencing.Conclusion The present PCR-TGGE method could be a useful tool for the molecular screening for the point mutations causing α-thalassemia and α-globin gene polymorphism.
