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. 1980 Dec 20;8(24):6007–6017. doi: 10.1093/nar/8.24.6007

The isolation of cloned cDNA sequences which are differentially expressed in human lymphocytes and fibroblasts.

J Crampton, S Humphries, D Woods, R Williamson
PMCID: PMC328068  PMID: 6162155

Abstract

Poly(A)+ RNA populations derived from normal lymphocytes and fibroblasts have been compared by hybridising each RNA to cDNA derived from the other RNA population. This indicated that approximately 75% of the sequences were common to both, and that these were present at different concentrations in the two cell types. The two RNA populations were further compared by hybridising them to a cDNA recombinant library derived from lymphocyte poly(A)+ RNA. This allowed the identification of clones containing sequences which are abundant in lymphocyte poly(A)+ RNA but absent or rare in fibroblast poly(A)+ RNA. A direct estimation of the abundance of five of these sequences in lymphocyte cDNA demonstrated that clones can be detected by such a procedure if they represent 0.2% or greater of the original cDNA population.

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Selected References

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

  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birnstiel M. L., Sells B. H., Purdom I. F. Kinetic complexity of RNA molecules. J Mol Biol. 1972 Jan 14;63(1):21–39. doi: 10.1016/0022-2836(72)90519-0. [DOI] [PubMed] [Google Scholar]
  3. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  5. Dworkin M. B., Dawid I. B. Use of a cloned library for the study of abundant poly(A)+RNA during Xenopus laevis development. Dev Biol. 1980 May;76(2):449–464. doi: 10.1016/0012-1606(80)90393-0. [DOI] [PubMed] [Google Scholar]
  6. Gergen J. P., Stern R. H., Wensink P. C. Filter replicas and permanent collections of recombinant DNA plasmids. Nucleic Acids Res. 1979 Dec 20;7(8):2115–2136. doi: 10.1093/nar/7.8.2115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Getz M. J., Birnie G. D., Young B. D., MacPhail E., Paul J. A kinetic estimation of base sequence complexity of nuclear poly(A)-containing RNA in mouse Friend cells. Cell. 1975 Feb;4(2):121–129. doi: 10.1016/0092-8674(75)90118-x. [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. Harris R., Ukaejiofo E. O. Tissue typing using a routine one-step lymphocyte separation procedure. Br J Haematol. 1970 Feb;18(2):229–235. doi: 10.1111/j.1365-2141.1970.tb01436.x. [DOI] [PubMed] [Google Scholar]
  10. Hastie N. D., Bishop J. O. The expression of three abundance classes of messenger RNA in mouse tissues. Cell. 1976 Dec;9(4 Pt 2):761–774. doi: 10.1016/0092-8674(76)90139-2. [DOI] [PubMed] [Google Scholar]
  11. Ostrow R. S., Woods W. G., Vosika G. J., Faras A. J. Analysis of the genetic complexity and abundance classes of messenger RNA in human liver and leukemic cells. Biochim Biophys Acta. 1979 Mar 28;562(1):92–102. doi: 10.1016/0005-2787(79)90129-1. [DOI] [PubMed] [Google Scholar]
  12. Radloff R., Bauer W., Vinograd J. A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells. Proc Natl Acad Sci U S A. 1967 May;57(5):1514–1521. doi: 10.1073/pnas.57.5.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Sim G. K., Kafatos F. C., Jones C. W., Koehler M. D., Efstratiadis A., Maniatis T. Use of a cDNA library for studies on evolution and developmental expression of the chorion multigene families. Cell. 1979 Dec;18(4):1303–1316. doi: 10.1016/0092-8674(79)90241-1. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Strohman R. C., Moss P. S., Micou-Eastwood J., Spector D., Przybyla A., Paterson B. Messenger RNA for myosin polypeptides: isolation from single myogenic cell cultures. Cell. 1977 Feb;10(2):265–273. doi: 10.1016/0092-8674(77)90220-3. [DOI] [PubMed] [Google Scholar]
  16. van de Sande J. H., Kleppe K., Khorana H. G. Reversal of bacteriophage T4 induced polynucleotide kinase action. Biochemistry. 1973 Dec 4;12(25):5050–5055. doi: 10.1021/bi00749a004. [DOI] [PubMed] [Google Scholar]

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