alpha-and beta-Globin complementary deoxyribonucleic acids of human and rabbit. Specificity of hybridization.
The specificity of hybridization was compared between the human and rabbit alpha and beta-globin complementary DNAs (cDNAs) and the corresponding alpha and beta-globin messenger RNAs (mRNAs). The globin chain-specific mRNAs of rabbit were prepared from polysomes incubated with O-methylthreonine (alpha and beta) or from postribosomal supernatant (alpha). Enrichment for either the alpha- or beta-globin mRNA was demonstrated by cell-free protein synthesis and by RNA-cDNA hybridization. Human mRNAs, active as templates for RNA-directed DNA polymerase, were prepared from reticulocytes of patients with hemolytic anemia, alpha-thalassemia (hemoglobin H disease), and beta-thalassemia. Because there was partial cross-hybridization between human mRNA and rabbit cDNA, the rabbit alpha- and beta-globin cDNAs could be used to demonstrate that the beta-thalassemia mRNA was enriched in human alpha-globin mRNA sequences and that the alpha-thalassemia mRNA was enriched in human beta-globin mRNA sequences. These results were confirmed by preparation of thalassemia globin cDNAs and subsequent hybridization to their template mRNAs. The amount of cross-hybridization between the human and rabbit alpha-globin mRNA and the two alpha-globin cDNAs was comparable to the cross-hybridization between the two beta-globin mRNAs and the two beta-globin cDNAs, indicating a similar degree of evolutionary divergence in the nucleotide sequences of the two globin genes.
D000818:Animals; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D004247:DNA / Q000097:blood*; D005914:Globins / Q000096:biosynthesis*; D006801:Humans; D009693:Nucleic Acid Hybridization; D011132:Polyribosomes / Q000032:analysis / Q000378:metabolism; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000097:blood; D011817:Rabbits; D012156:Reticulocytes / Q000032:analysis / Q000378:metabolism; D013045:Species Specificity; D014158:Transcription, Genetic
D012333:RNA, Messenger; D005914:Globins; D004247:DNA