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Synthesis of Oligonucleotides Used as Probes; Purification by HPLC

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Peroxisomes

Part of the book series: Springer Laboratory ((SLM))

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Abstract

Synthetic oligonucleotides are useful research tools in molecular biology [3, 7]. As hybridization probes, they can distinguish a perfect match in the target sequences from various single-base-pair mismatches [20, 21]. They have been used to introduce mutations in DNA at specific sites [8, 13, 15, 22, 23] as well as to probe for mutations at specific loci in genes [4, 9, 13, 20, 22, 23]. They have also been employed in the isolation of specific DNA sequences from a bank of cloned cDNA [6, 16, 19, 21] and in the determination of RNA secondary structure [11]. Moreover, the use of antisense DNA or RNA oligonucleotides to block the translation or processing of specific mRNAs provides a powerful tool for studies of the regulation of gene expression and development [2, 5].

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References

  1. Causeret C, Bentéjac M, Bugaut M (1993) Proteins and enzymes of the peroxisomal membrane in mammals. Biol Cell 77:89–104

    Article  PubMed  CAS  Google Scholar 

  2. Cohen JS (1991) Antisense oligodeoxynucleotides as antiviral agents. Antiviral Res 16:121–133

    Article  PubMed  CAS  Google Scholar 

  3. Eckstein F (1991) Oligonucleotides and analogues: A practical approach, IRL Press, Oxford

    Google Scholar 

  4. Farr CJ, Saiki RK, Erlich HA, McCormick F, Marshall CJ (1988) Analysis of RAS gene mutations in acute myeloid leukemia by polymerase chain reaction and oligonucleotide probes. Proc Natl Acad Sci USA 85:1629–1633

    Article  PubMed  CAS  Google Scholar 

  5. Hélène C, Toulmé JJ (1990) Specific regulation of gene expression by antisense, sense and antigene nucleic acids. Biochim Biophys Acta 1049:99–125

    Article  PubMed  Google Scholar 

  6. Heller RA, Song K, Freire-Moar J (1992) Rapid screening of libraries with radiolabeled DNA sequences generated by PCR using highly degenerate oligonucleotide mixtures. Biotechniques 12:30–35

    PubMed  CAS  Google Scholar 

  7. Itakura K, Rossi JJ, Wallace RB (1984) Synthesis and use of synthetic oligonucleotides. Annu Rev Biochem 53:323–356

    Article  PubMed  CAS  Google Scholar 

  8. Ito W, Ishiguro H, Kurosawa Y (1991) A general method for introducing a series of mutations into cloned DNA using the polymerase chain reaction. Gene 102:67–70

    Article  PubMed  CAS  Google Scholar 

  9. Karnik SS, Khorana GH (1990) Assembly of functional rhodopsin requires a disulfure bond between cysteine residues 110 and 187. J Biol Chem 265:17520–17524

    PubMed  CAS  Google Scholar 

  10. Latruffe N (1992) Les peroxysomes et la proliferation cellulaire ou la prise en considération d’un organite méconnu. Médecine/Sciences 8:239–248

    Google Scholar 

  11. McWilliams RA, Glitz DG (1991) Localization of a segment of 16S RNA on the surface of the small ribosomal subunit by immune electron microscopy of complementary oligodeoxynucleotides. Biochimie 73:911–918

    Article  PubMed  CAS  Google Scholar 

  12. Miyazawa S, Hayashi H, Hijikata M, Ishii N, Furuta S, Kagamiyamla H, Osumi T, Hashimoto T (1987) Complete nucleotide sequence of cDNA and predicted aminoacide sequence of rat acyl-CoA oxidase. J Biol Chem 262:8131–8137

    PubMed  CAS  Google Scholar 

  13. Norrander J, Kempe T, Messing J (1983) Construction of improved M13 vectors using oligodeoxynucleo tide-directed mutagenesis. Gene 26:101–106

    Article  PubMed  CAS  Google Scholar 

  14. Osumi T, Ozasa H, Miyazawa S, Hashimoto T (1984) Molecular cloning of cDNA for rat liver catalase. Biochem Biophys Res Commun 122:831–837

    Article  PubMed  CAS  Google Scholar 

  15. Sayers JR, Krekel C, Eckstein F (1992) Rapid high-efficiency site-directed mutagenesis by the phosphorothioate approach. Biotechniques 13:592–596

    PubMed  CAS  Google Scholar 

  16. Suggs SV, Wallace BR, Hirose T, Kawashima EH, Itakura K (1981) Use of synthetic oligonucleotides as hybridization probes: isolation of cloned cDNA sequences for human β2-microglobulin. Proc Natl Acad Sci USA 78:6613–6617

    Article  PubMed  CAS  Google Scholar 

  17. Suzuki H, Kawarabayasi Y, Kondo J, Abe T, Nishikawa K, Kimura S, Hashimoto T, Yamamoto T (1990) Structure and regulation of rat long-chain acyl-CoA synthetase. J Biol Chem 265:8681–8685

    PubMed  CAS  Google Scholar 

  18. Van Den Bosch H, Schutgens RBH, Wanders RJA, Tager JM (1992) Biochemistry of peroxisomes. Annu Rev Biochem 61:157–197

    Article  Google Scholar 

  19. Wallace RB, Johnson MJ, Hirose T, Miyake T, Kawashima EH, Itakura K (1981) The use of synthetic oligonucleotides as hybridization probes. II. Hybridization of oligonucleotides of mixed sequence to rabbit β-globin DNA. Nucl Acids Res 9:879–894

    Article  PubMed  CAS  Google Scholar 

  20. Wallace RB, Shaffer J, Bonner J, Hirose T, Itakura K (1979) Hybridization of synthetic oligodeoxyribonucleotides to φ χ 174 DNA: the effect of single base pair mismatch. Nucl Acids Res 6:3543–3557

    Article  PubMed  CAS  Google Scholar 

  21. Wood WI, Gitschier J, Lasky LA, Lawn RM (1985) Base composition-independent hybridization in tetramethylammonium chloride: A method for oligonucleotide screening of highly complex gene libraries. Proc Natl Acad Sci USA 82:1585–1588

    Article  PubMed  CAS  Google Scholar 

  22. Zoller MJ, Smith M (1982) Oligonucleotide-directed mutagenesis using M13-derived vectors: an efficient and general procedure for the production of point mutations in any fragment of DNA. Nucl Acids Res 10:6487–6500

    Article  PubMed  CAS  Google Scholar 

  23. Zoller MJ, Smith M (1983) Oligonucleotide-directed mutagenesis of DNA fragments cloned into M13 vectors. In Wu R, Grossman L and Moldave K (eds). Methods in Enzymology, Vol. 100, Academic, New York, pp. 468–500

    Google Scholar 

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© 1994 Springer-Verlag Berlin Heidelberg

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Bentéjac, M., Bugaut, M. (1994). Synthesis of Oligonucleotides Used as Probes; Purification by HPLC. In: Latruffe, N., Bugaut, M. (eds) Peroxisomes. Springer Laboratory. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-87807-7_7

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  • DOI: https://doi.org/10.1007/978-3-642-87807-7_7

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-87809-1

  • Online ISBN: 978-3-642-87807-7

  • eBook Packages: Springer Book Archive

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