Organization of human δ- and β-globin genes in cellular DNA and the presence of intragenic inserts

Localization of the site of recombination in formation of the Lepore Boston globin gene.

“…and Asu I (Bio-Lab Inc., Decatur, Ga.) was performed according to the manufacturer's specifications. Transfer of DNA fragments from 0.8% agarose (Sea Kem) to nitrocellulose filters, nick translation of probes (1-8 x 108 cpm/,g), hybridization of filters, and radioautography were performed as described previously (2,9).…”

Heterogeneity of DNA fragments associated with the sickle-globin gene.

“…Newer techniques, including the use of restriction endonucleases, now permit an analysis of such polymorphisms at the gene level by detecting changes in DNA sequence organization (1). Restriction endonuclease mapping with the Southern "blotting" technique has provided detailed information on the organization of the human 6-and f3-globin structural genes (2)(3)(4)(5). This procedure involves the cleavage of cellular DNA with restriction Received for publication 6 April 1979 and in revised form 14 May 1979. enzymes, separation ofthe DNA fragments by size with agarose gel electrophoresis, transfer of the DNA to nitrocelltilose filters, and subsequent hybridization to specific radioactive globin complementary DNA (cDNA)1 probes.…”

“…The cDNA probe was nick-translated, as described, to a 2-4 x 108 cpm/,ug sp act (8). The hydbridization, washing of filters, and autoradiography were performed as described (4,5). A Cot value of 0.001 was attained after hybridization at 68°C for 24 h (4, 5).…”

The Structure and Expression of Globin Genes in Rabbit and Man

“…between the codons for amino acids 101-120. The structure of the 6-3-gene locus has also been elucidated by Mears et al [26] and Lawn et al [27]. In the latter case [27], the linkage of the 6-and 3-globin genes has also been demonstrated.…”