Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1985 Nov;82(22):7510–7514. doi: 10.1073/pnas.82.22.7510

Modified oligonucleotides as alternatives to the synthesis of mixed probes for the screening of cDNA libraries.

T Huynh-Dinh, N Duchange, M M Zakin, A Lemarchand, J Igolen
PMCID: PMC390846  PMID: 3865174

Abstract

Two simple alternatives to the synthesis of mixed oligodeoxyribonucleotide probes are described: mixed deblocking of triazolodeoxynucleosides for the A/G and C/T degeneracies or incorporation of 2-amino-2'-deoxyadenosines into a determined sequence for a higher stability of the hybridization duplexes. The synthetic oligodeoxyribonucleotides obtained were successfully tested in comparison with a classical mixed probe for their capacity to identify specific cDNA clones of human antithrombin III. The results are discussed with respect to the utilization of these synthetic oligonucleotides as hybridization probes for the isolation of cloned cDNA sequences.

Full text

PDF
7512

Images in this article

7512Image
on p.7512
7512Image
on p.7512
7512Image
on p.7512
7512Image
on p.7512
7512Image
on p.7512
7512Image
on p.7512
7512Image
on p.7512
7512Image
on p.7512
7512Image
on p.7512
7512Image
on p.7512
7513Image
on p.7513
7513Image
on p.7513
7513Image
on p.7513
7513Image
on p.7513
7513Image
on p.7513
7513Image
on p.7513

Selected References

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

  1. Anderson S., Kingston I. B. Isolation of a genomic clone for bovine pancreatic trypsin inhibitor by using a unique-sequence synthetic DNA probe. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6838–6842. doi: 10.1073/pnas.80.22.6838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anson D. S., Choo K. H., Rees D. J., Giannelli F., Gould K., Huddleston J. A., Brownlee G. G. The gene structure of human anti-haemophilic factor IX. EMBO J. 1984 May;3(5):1053–1060. doi: 10.1002/j.1460-2075.1984.tb01926.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bock S. C., Wion K. L., Vehar G. A., Lawn R. M. Cloning and expression of the cDNA for human antithrombin III. Nucleic Acids Res. 1982 Dec 20;10(24):8113–8125. doi: 10.1093/nar/10.24.8113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bresser J., Doering J., Gillespie D. Quick-blot: selective mRNA or DNA immobilization from whole cells. DNA. 1983;2(3):243–254. doi: 10.1089/dna.1983.2.243. [DOI] [PubMed] [Google Scholar]
  5. Chandra T., Stackhouse R., Kidd V. J., Woo S. L. Isolation and sequence characterization of a cDNA clone of human antithrombin III. Proc Natl Acad Sci U S A. 1983 Apr;80(7):1845–1848. doi: 10.1073/pnas.80.7.1845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fisher E. F., Caruthers M. H. Color coded triarylmethyl protecting groups useful for deoxypolynucleotide synthesis. Nucleic Acids Res. 1983 Mar 11;11(5):1589–1599. doi: 10.1093/nar/11.5.1589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gaffney B. L., Marky L. A., Jones R. A. The influence of the purine 2-amino group on DNA conformation and stability. Synthesis and conformational analysis of d[T(2-aminoA)]3. Nucleic Acids Res. 1982 Jul 24;10(14):4351–4361. doi: 10.1093/nar/10.14.4351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Kalderon D., Richardson W. D., Markham A. F., Smith A. E. Sequence requirements for nuclear location of simian virus 40 large-T antigen. Nature. 1984 Sep 6;311(5981):33–38. doi: 10.1038/311033a0. [DOI] [PubMed] [Google Scholar]
  10. Millican T. A., Mock G. A., Chauncey M. A., Patel T. P., Eaton M. A., Gunning J., Cutbush S. D., Neidle S., Mann J. Synthesis and biophysical studies of short oligodeoxynucleotides with novel modifications: a possible approach to the problem of mixed base oligodeoxynucleotide synthesis. Nucleic Acids Res. 1984 Oct 11;12(19):7435–7453. doi: 10.1093/nar/12.19.7435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Park I., Schaeffer E., Sidoli A., Baralle F. E., Cohen G. N., Zakin M. M. Organization of the human transferrin gene: direct evidence that it originated by gene duplication. Proc Natl Acad Sci U S A. 1985 May;82(10):3149–3153. doi: 10.1073/pnas.82.10.3149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Prochownik E. V., Markham A. F., Orkin S. H. Isolation of a cDNA clone for human antithrombin III. J Biol Chem. 1983 Jul 10;258(13):8389–8394. [PubMed] [Google Scholar]
  13. Sharpe C. R., Sidoli A., Shelley C. S., Lucero M. A., Shoulders C. C., Baralle F. E. Human apolipoproteins AI, AII, CII and CIII. cDNA sequences and mRNA abundance. Nucleic Acids Res. 1984 May 11;12(9):3917–3932. doi: 10.1093/nar/12.9.3917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sung W. L. Synthesis of 4-triazolopyrimidinone nucleotide and its application in synthesis of 5-methylcytosine-containing oligodeoxyribonucleotides. Nucleic Acids Res. 1981 Nov 25;9(22):6139–6151. doi: 10.1093/nar/9.22.6139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Taboury J. A., Adam S., Taillandier E., Neumann J. M., Tran-Dinh S., Huynh-Dinh T., Langlois d'Estaintot B., Conti M., Igolen J. The B----Z transition in two synthetic oligonucleotides: d(C-2-amino-ACGTG) and d(m5CGCAm5CGTGCG) studied by IR, NMR and CD spectroscopies. Nucleic Acids Res. 1984 Aug 10;12(15):6291–6305. doi: 10.1093/nar/12.15.6291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Wallace R. B., Johnson M. J., Hirose T., Miyake T., Kawashima E. H., Itakura K. The use of synthetic oligonucleotides as hybridization probes. II. Hybridization of oligonucleotides of mixed sequence to rabbit beta-globin DNA. Nucleic Acids Res. 1981 Feb 25;9(4):879–894. doi: 10.1093/nar/9.4.879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wallace R. B., Shaffer J., Murphy R. F., Bonner J., Hirose T., Itakura K. Hybridization of synthetic oligodeoxyribonucleotides to phi chi 174 DNA: the effect of single base pair mismatch. Nucleic Acids Res. 1979 Aug 10;6(11):3543–3557. doi: 10.1093/nar/6.11.3543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Whitehead A. S., Goldberger G., Woods D. E., Markham A. F., Colten H. R. Use of a cDNA clone for the fourth component of human complement (C4) for analysis of a genetic deficiency of C4 in guinea pig. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5387–5391. doi: 10.1073/pnas.80.17.5387. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES