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Title: Relative quantitation of mRNA in β-thalassemia/Hb E using real-time polymerase chain reaction
Authors: Yuwadee Watanapokasin
P. Winichagoon
S. Fuchareon
P. Wilairat
Srinakharinwirot University
The Institute of Science and Technology for Research and Development, Mahidol University
Mahidol University
Keywords: Biochemistry, Genetics and Molecular Biology;Medicine
Issue Date: 1-Jan-2000
Citation: Hemoglobin. Vol.24, No.2 (2000), 105-116
Abstract: β-Thalassemia and Hb E patients, with seemingly identical genotypes, have a remarkable variability in severity. Reduction in red cell survival in β-thalassemia is correlated with the amount of intracellular unmatched α- globin chains. However, it was only recently realized that mRNA, whose translation is prematurely terminated, is also unstable. No systematic attempts have been made to investigate mRNA stability in β-thalassemia arising from nonsense mutations located upstream from the normal termination codon. In this study, one-step real-time polymerase chain reaction has been employed to compare the levels of α- and β-globin mRNA in reticulocytes from β-thalassemia/Hb E subjects. The results showed the highest α/β- globin mRNA ratio (median = 5.70, n = 13) in frameshift codons 41/42 (- TTCT)/Hb E individuals compared to normal subjects (median = 1.02, n = 6), or those with Hb E trait (median = 2.15, n = 8). In addition, there was a concomitant increase in the α/β-globin mRNA ratio with decrease in hemoglobin level, i.e., increase in severity. The difference in the ratio among β-thalassemia/Hb E patients with the same genotype may be attributed to individual variations of efficiency in β(E)-globin mRNA splicing and in the destruction of prematurely terminated mRNA.
ISSN: 03630269
Appears in Collections:Scopus 1991-2000

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