期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1973
卷号：70
期号：6
页码：1886-1890
DOI：10.1073/pnas.70.6.1886
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：In previous studies of patients with {beta} thalassemia, mRNA extracted from reticulocytes in peripheral blood when added to cell-free systems reproduces the deficient {beta}-chain synthesis characteristic of intact cells. The present studies with specific probes for [α] and {beta} mRNA were designed to decide whether the decreased {beta} mRNA activity is due to the presence of abnormal or reduced {beta} globin mRNA in these cells. Purified [α] and {beta} complementary DNAs (cDNAs) have been synthesized with RNA-instructed DNA polymerase; [α] and {beta} mRNAs isolated from heavy ({beta}-producing) and light ([α]-producing) polyribosomes of rabbit reticulocytes were used as templates. Each of the cDNAs is more than 80% pure by the criterion of biological activity. The [α] cDNA labeled with [32P]dCTP and the {beta} cDNA labeled with [3H]dCTP have been added simultaneously to reaction mixtures containing various concentrations of mRNA from thalassemic and nonthalassemic subjects. The extent and rate of hybridization were determined, permitting a comparison of relative [α] and {beta} mRNA content in the same annealing mixture. In six nonthalassemic patients, relatively equal amounts of hybridizable [α] and {beta} mRNA appear to be present. In five of seven patients with {beta}-thalassemia, significantly decreased amounts of {beta} mRNA compared to [α] mRNA can be demonstrated. In two patients with Hemoglobin H disease, there is a decreased amount of [α] mRNA compared to {beta} mRNA.
