Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1978 Feb;5(2):563–581. doi: 10.1093/nar/5.2.563

Insertion of synthetic copies of human globin genes into bacterial plasmids.

J T Wilson, L B Wilson, J K deRiel, L Villa-komaroff, A Efstratiadis, B G Forget, S M Weissman
PMCID: PMC342002  PMID: 345245

Abstract

Double stranded human globin cDNA was synthesized by use of viral reverse transcriptase from globin mRNA of cord blood of premature infants requiring exchange transfusions. The cDNA was introduced into plasmids and the recombinant DNA plasmids used to transform E. coli X1776. A number of transformants were obtained. Plasmid DNA from selected colonies was isolated and characterized for the type of globin cDNA it contained by three types of procedures: 1) hybridization to previously characterized 3H-labeled alpha,beta and gamma cDNA; 2) analysis of the size and nature of fragments produced by digestion of the plasma DNA by different restriction endonucleases; and 3) by rapid DNA sequence analysis of selected DNA fragments produced by restriction endonuclease digestion. Analysis by these techniques of plasmid DNA from different colonies has definitively identified the presence of human alpha, beta or gamma cDNA sequences in different plasmids.

Full text

PDF
563

Images in this article

569Image
on p.569
572Image
on p.572
573Image
on p.573
575Image
on p.575
577Image
on p.577
578Image
on p.578

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Benz E. J., Jr, Swerdlow P. S., Forget B. G. Absence of functional messenger RNA activity for beta globin chain synthesis in beta 0-thalassemia. Blood. 1975 Jan;45(1):1–10. [PubMed] [Google Scholar]
  2. Chang J. C., Temple G. F., Poon R., Neumann K. H., Kan Y. W. The nucleotide sequences of the untranslated 5' regions of human alpha- and beta-globin mRNAs. Proc Natl Acad Sci U S A. 1977 Nov;74(11):5145–5149. doi: 10.1073/pnas.74.11.5145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cohen-Solal M., Forget B. G., Prensky W., Marotta C. A., Weissman S. M. Human beta-globin messenger RNA. II. nucleotide sequences derived from 125I-labeled globin messenger RNA. J Biol Chem. 1977 Jul 25;252(14):5032–5039. [PubMed] [Google Scholar]
  4. Covey C., Richardson D., Carbon J. A method for the deletion of restriction sites in bacterial plasmid deoxyribonucleic acid. Mol Gen Genet. 1976 May 7;145(2):155–158. doi: 10.1007/BF00269587. [DOI] [PubMed] [Google Scholar]
  5. Efstratiadis A., Kafatos F. C., Maxam A. M., Maniatis T. Enzymatic in vitro synthesis of globin genes. Cell. 1976 Feb;7(2):279–288. doi: 10.1016/0092-8674(76)90027-1. [DOI] [PubMed] [Google Scholar]
  6. Efstratiadis A., Maniatis T., Kafatos F. C., Jeffrey A., Vournakis J. N. Full length and discrete partial reverse transcripts of globin and chorion mRNAs. Cell. 1975 Apr;4(4):367–378. doi: 10.1016/0092-8674(75)90157-9. [DOI] [PubMed] [Google Scholar]
  7. Forget B. G., Hillman D. G., Lazarus H., Barell E. F., Benz ej J. R., Caskey C. T., Huisman T. H., Schroeder W. A., Housman D. Absence of messenger RNA and gene DNA for beta-globin chains in hereditary persistence of fetal hemoglobin. Cell. 1976 Mar;7(3):323–329. doi: 10.1016/0092-8674(76)90161-6. [DOI] [PubMed] [Google Scholar]
  8. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Guerry P., LeBlanc D. J., Falkow S. General method for the isolation of plasmid deoxyribonucleic acid. J Bacteriol. 1973 Nov;116(2):1064–1066. doi: 10.1128/jb.116.2.1064-1066.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Higuchi R., Paddock G. V., Wall R., Salser W. A general method for cloning eukaryotic structural gene sequences. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3146–3150. doi: 10.1073/pnas.73.9.3146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Housman D., Forget B. G., Skoultchi A., Benz E. J., Jr Quantitative deficiency of chain-specific globin messenger ribonucleic acids in the thalassemia syndromes. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1809–1813. doi: 10.1073/pnas.70.6.1809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kazazian H. H., Jr, Ginder G. D., Snyder P. G., Van Beneden R. J., Woodhead A. P. Further evidence of a quantitative deficiency of chain-specific globin mRNA in the thalassemia syndromes. Proc Natl Acad Sci U S A. 1975 Feb;72(2):567–571. doi: 10.1073/pnas.72.2.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lanyon W. G., Ottolenghi S., Williamson R. Human globin gene expression and linkage in bone marrow and fetal liver. Proc Natl Acad Sci U S A. 1975 Jan;72(1):258–262. doi: 10.1073/pnas.72.1.258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Maizels N. Dictyostelium 17S, 25S, and 5S rDNAs lie within a 38,000 base pair repeated unit. Cell. 1976 Nov;9(3):431–438. doi: 10.1016/0092-8674(76)90088-x. [DOI] [PubMed] [Google Scholar]
  15. Maniatis T., Kee S. G., Efstratiadis A., Kafatos F. C. Amplification and characterization of a beta-globin gene synthesized in vitro. Cell. 1976 Jun;8(2):163–182. doi: 10.1016/0092-8674(76)90001-5. [DOI] [PubMed] [Google Scholar]
  16. Marotta C. A., Wilson J. T., Forget B. G., Weissman S. M. Human beta-globin messenger RNA. III. Nucleotide sequences derived from complementary DNA. J Biol Chem. 1977 Jul 25;252(14):5040–5053. [PubMed] [Google Scholar]
  17. Proudfoot N. J., Longley J. I. The 3' terminal sequences of human alpha and beta globin messenger RNAs: comparison with rabbit globin messenger RNA. Cell. 1976 Dec;9(4 Pt 2):733–746. doi: 10.1016/0092-8674(76)90137-9. [DOI] [PubMed] [Google Scholar]
  18. Roychoudhury R., Jay E., Wu R. Terminal labeling and addition of homopolymer tracts to duplex DNA fragments by terminal deoxynucleotidyl transferase. Nucleic Acids Res. 1976 Jan;3(1):101–116. doi: 10.1093/nar/3.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  20. Subramanian K. N., Pan J., Zain S., Weissman S. M. The mapping and ordering of fragments of SV40 DNA produced by restriction endonucleases. Nucleic Acids Res. 1974 Jun;1(6):727–752. doi: 10.1093/nar/1.6.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wilson J. T., Forget B. G., Wilson L. B., Weissman S. M. Human globin messenger RNA: importance of cloning for structural analysis. Science. 1977 Apr 8;196(4286):200–202. doi: 10.1126/science.847468. [DOI] [PubMed] [Google Scholar]
  22. Wilson J. T., deRiel J. K., Forget B. G., Marotta C. A., Weissman S. M. Nucleotide sequence of 3' untranslated portion of human alpha globin mRNA. Nucleic Acids Res. 1977 Jul;4(7):2353–2368. doi: 10.1093/nar/4.7.2353. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES