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The Isolation and Characterization of Linked δ- and β-Globin Genes from a Cloned Library of Human DNA

Lawn, Richard M. and Fritsch, Edward F. and Parker, Richard C. and Blake, Geoffrey and Maniatis, Tom (1978) The Isolation and Characterization of Linked δ- and β-Globin Genes from a Cloned Library of Human DNA. Cell, 15 (4). pp. 1157-1174. ISSN 0092-8674. doi:10.1016/0092-8674(78)90043-0.

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A cloned library of large, random embryonic human DNA fragments was constructed and screened for β-globin sequences using the cloned human β-globin cDNA plasmid pJW102 (Wilson et al., 1978) as a hybridization probe. Two independent clones were obtained and then characterized by restriction endonuclease cleavage analysis, hybridization experiments and partial DNA sequencing. Each of the clones carries both the adult δ- and β-globin genes. The two genes are separated by approximately 5.4 kilobases (kb) of DNA and their orientation with respect to the direction of transcription is 5′-δ-β-3′. Both the δ-and β-globin genes contain a large noncoding intervening sequence (950 and 900 bp, respectively) located between the codons for amino acids 104 (arginine) and 105 (leucine). Although the location of the large intervening sequence within the coding regions of the two genes is identical, the two noncoding sequences bear little sequence homology. A second, smaller intervening sequence similar to that found in other mammalian β-globin genes was detected near the 5′ end of the human β-globin gene. The two independently isolated β-globin clones differ from each other by the presence of a Pst I restriction enzyme cleavage site within the large intervening sequence of the δ-globin gene of one of the clones. This suggests that the human DNA carried in the two clones was derived from two homologous chromosomes which were heterozygous for the Pst I restriction enzyme recognition sequence.

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Additional Information:© 1978 Elsevier Inc. Received August 28, 1978. We are grateful to Bernard Forget for advice and discussions, and for providing fetal liver DNA. We thank J. Wilson, L. Wilson, B. Forget and S. Weissman for providing human globin cDNA clones, and C. O'Connell for help in constructing and screening the human DNA library. We also thank J. Posakony for help with the DNA sequencing, J. Maniatis for assistance in preparing graphics and A. Cortenbach for preparing media and materials. This work was supported by a grant from the NIH and by funds from a Biomedical Research Support grant to the California Institute of Technology. R.M.L. and E.F.F. were supported by postdoctoral fellowships from the American Cancer Society and the Damon Runyon-Walter Winchell Cancer Fund, respectively. T.M. is a recipient of a Rita Allen Foundation career development award. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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Biomedical Research Support GrantUNSPECIFIED
American Cancer SocietyUNSPECIFIED
Damon Runyon-Walter Winchell Cancer FundUNSPECIFIED
Rita Allen FoundationUNSPECIFIED
Issue or Number:4
Record Number:CaltechAUTHORS:20120911-143053154
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:34005
Deposited By: Ruth Sustaita
Deposited On:11 Sep 2012 22:10
Last Modified:09 Nov 2021 23:05

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