Abstract
A rapid method for the synthesis of oligodeoxynucleotides (ODNs) terminated by 5'-amino-5'-deoxythymidine is described. A 3'-phosphorylated ODN (the donor) is incubated in aqueous solution with 5'-amino- 5'-deoxythymidine in the presence of N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC), extending the donor by one residue via a phosphoramidate bond. Template- directed ligation of the extended donor and an acceptor ODN, followed by acid hydrolysis, yields the acceptor ODN extended by a single 5'-amino-5'-deoxythymidine residue at its 5'terminus.
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- Bruick R. K., Dawson P. E., Kent S. B., Usman N., Joyce G. F. Template-directed ligation of peptides to oligonucleotides. Chem Biol. 1996 Jan;3(1):49–56. doi: 10.1016/s1074-5521(96)90084-8. [DOI] [PubMed] [Google Scholar]
- Chu B. C., Wahl G. M., Orgel L. E. Derivatization of unprotected polynucleotides. Nucleic Acids Res. 1983 Sep 24;11(18):6513–6529. doi: 10.1093/nar/11.18.6513. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Letsinger F. L., Wilkes J. S. Incorporation of 5'-amino-5'-deoxythymidine5'-phosphate in polynucleotides by use of DNA polymerase I and a phiX174 DNA template. Biochemistry. 1976 Jun 29;15(13):2810–2816. doi: 10.1021/bi00658a017. [DOI] [PubMed] [Google Scholar]
- Letsinger R. L., Mungall W. S. Phosphoramidate analogs of oligonucleotides. J Org Chem. 1970 Nov;35(11):3800–3803. doi: 10.1021/jo00836a048. [DOI] [PubMed] [Google Scholar]
- Smith L. M., Fung S., Hunkapiller M. W., Hunkapiller T. J., Hood L. E. The synthesis of oligonucleotides containing an aliphatic amino group at the 5' terminus: synthesis of fluorescent DNA primers for use in DNA sequence analysis. Nucleic Acids Res. 1985 Apr 11;13(7):2399–2412. doi: 10.1093/nar/13.7.2399. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith L. M., Kaiser R. J., Sanders J. Z., Hood L. E. The synthesis and use of fluorescent oligonucleotides in DNA sequence analysis. Methods Enzymol. 1987;155:260–301. doi: 10.1016/0076-6879(87)55021-2. [DOI] [PubMed] [Google Scholar]
- Sproat B. S., Beijer B., Rider P. The synthesis of protected 5'-amino-2',5'-dideoxyribonucleoside-3'-O-phosphoramidites; applications of 5'-amino-oligodeoxyribonucleotides. Nucleic Acids Res. 1987 Aug 11;15(15):6181–6196. doi: 10.1093/nar/15.15.6181. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tung C. H., Rudolph M. J., Stein S. Preparation of oligonucleotide-peptide conjugates. Bioconjug Chem. 1991 Nov-Dec;2(6):464–465. doi: 10.1021/bc00012a015. [DOI] [PubMed] [Google Scholar]