Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1976 Apr;3(4):1053–1063. doi: 10.1093/nar/3.4.1053

Enzymatic synthesis of duplex circular phiX174 DNA containing phosphoramidate bonds in the (-) strand.

R L Letsinger, B Hapke, G R Petersen, L B Dumas
PMCID: PMC342964  PMID: 775443

Abstract

Duplex circular phiX174 DNA (RF I) containing some phosphoramidate links in the backbone chain of the (-) strand was synthesized by reaction of 5'-amino-5'-deoxythymidine 5'-triphosphate, dCTP, dGTP, and 3H-dATP with DNA polymerase I and DNA ligase (T4) on a (+) strand phiX174 amber 3 DNA template. The yield of duplex DNA was higher when dTTP was included along with the amino analog in the initial reaction system or was added late in the synthesis. RF I DNA was observed as a rapidly sedimenting species in an alkaline sucrose gradient, and the presence of phosphoramidate linkages was demonstrated by the unusual lability of the duplex DNA in a weakly acidic solution.

Full text

PDF
1053

Selected References

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

  1. Dumas L. B., Darby G., Sinsheimer R. L. The replication of bacteriophage phi X174 DNA in vitro. Temperature effects on repair synthesis and displacement synthesis. Biochim Biophys Acta. 1971 Jan 28;228(2):407–422. [PubMed] [Google Scholar]
  2. GUTHRIE G. D., SINSHEIMER R. L. Observations on the infection of bacterial protoplasts with the deoxyribonucleic acid of bacteriophage phi X174. Biochim Biophys Acta. 1963 Jun 25;72:290–297. [PubMed] [Google Scholar]
  3. Goulian M. Incorporation of oligodeoxynucleotides into DNA. Proc Natl Acad Sci U S A. 1968 Sep;61(1):284–291. doi: 10.1073/pnas.61.1.284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. JOSSE J., KORNBERG A. Glucosylation of deoxyribonucleic acid. III. alpha- and beta-Glucosyl transferases from T4-infected Escherichia coli. J Biol Chem. 1962 Jun;237:1968–1976. [PubMed] [Google Scholar]
  5. 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]
  6. Letsinger R. L., Wilkes J. S., Dumas L. B. Enzymatic synthesis of polydeoxyribonucleotides possessing internucleotide phosphoramidate bonds. J Am Chem Soc. 1972 Jan 12;94(1):292–293. doi: 10.1021/ja00756a060. [DOI] [PubMed] [Google Scholar]

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

RESOURCES