Human globin gene analysis for a patient with beta-o/delta beta-thalassemia.
Complementary DNA (cDNA) was prepared with RNA-dependent DNA polymerase from human globin messenger RNA (mRNA). Annealing and translation experimenta with total mRNA from circulating cells from a patient with heterozygous beta/heterozygous beta-delta-o thalassemia (beta-o/delta beta-o-thalassemia) demonstrated no detectable mRNA for beta-globin. cDNA enriched in sequences homologous to beta-globin mRNA was prepared by hydroxylapatite fractionation of hybrids formed between beta-o/delta beta-o-thalassemic mRNA and cDNA made from mRNA from a patient with alpha-thalassemia (hemoglobin H disease). The rate of annealing of this beta-enriched cDNA to normal human nuclear DNA was that of a sequence present as only a single copy per haploid genome. The beta-enriched cDNA annealed to the beta-o-delta beta-o-thalassemia total DNA with approximately the same kinetics as to normal DNA, indicating that no total gene deletion of beta-globin genes from the diploid genome has occurred, although the accuracy of the technique could not exclude with certainty a partial deletion or a deletion of a beta-globin gene from only one of the haploid genomes. This demonstrates that at least one of the beta-o- or the delta beta-o-thalassemia haploid genomes in this case contains a substantially intact beta-globin gene.
D000293:Adolescent; D009189:Avian Myeloblastosis Virus / Q000201:enzymology; D002845:Chromatography; D004247:DNA; D005260:Female; D005796:Genes; D005826:Genetics, Medical; D005914:Globins / Q000096:biosynthesis; D006579:Heterozygote; D006801:Humans; D006882:Hydroxyapatites; D008297:Male; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000097:blood; D012194:RNA-Directed DNA Polymerase; D012156:Reticulocytes / Q000378:metabolism; D013789:Thalassemia / Q000097:blood* / Q000378:metabolism
D006882:Hydroxyapatites; D012333:RNA, Messenger; D005914:Globins; D004247:DNA; D012194:RNA-Directed DNA Polymerase