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. 1978 Mar;75(3):1222–1226. doi: 10.1073/pnas.75.3.1222

Changes in restricted human cellular DNA fragments containing globin gene sequences in thalassemias and related disorders

J Gregory Mears *,†,‡, Francesco Ramirez *,†,‡, David Leibowitz *,†,‡, Frank Nakamura *,†,‡, Arthur Bloom *,†,‡, Felix Konotey-Ahulu †, Arthur Bank *,†,‡
PMCID: PMC411442  PMID: 274714

Abstract

Human cellular DNA fragments from cells of normal subjects and patients with thalassemia obtained by restriction enzyme digestion were analyzed for their globin gene content. The fragments were separated on agarose gels, transferred to nitrocellulose filters, hybridized to globin [32P]cDNA, and radioautographed. One to ten picograms of globin gene sequences were detectable. With EcoRI digestion, eight to nine cellular DNA fragments were found to contain globin genes. Three of these contained β-like gene sequences assayed with β globin cDNA probe. One β-like fragment was absent in DNA from a homozygous subject for hemoglobin Lepore. Two of the three β gene-containing fragments present in normal DNA were absent in DNA from a patient with hereditary persistence of fetal hemoglobin. The same two fragments containing β-like genes were absent from δβ thalassemic DNA and one new fragment containing β-like genes was found. Together with results obtained by hybridization of these DNAs in solution, the data are consistent with deletion of specific restriction human DNA fragments in subjects with these disorders and a greater deletion of β-like gene sequences in subjects with hereditary persistence of fetal hemoglobin than in those with δβ thalassemia.

Keywords: β thalassemia, restriction enzyme digestion, “blot” hybridization, cellular DNA

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