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. 1982 Aug;79(15):4594–4598. doi: 10.1073/pnas.79.15.4594

Site-directed psoralen crosslinking of DNA.

W A Saffran, M Goldenberg, C R Cantor
PMCID: PMC346721  PMID: 6289307

Abstract

A technique has been developed for placing site-specific crosslinks into DNA by using a psoralen derivative containing a sulfhydryl group. Plasmid pBR322 was nicked at the unique BamHI restriction site, and mercurated pyrimidines were introduced by nick-translation. Then the psoralen was specifically directed to this site in the dark through a Hg-S linkage prior to irradiation to form a crosslink. The location of interstrand cross-links was examined by electron microscopy of Pst I-cut molecules and found to be at or near the BamHI site.

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Selected References

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

  1. Bolivar F., Backman K. Plasmids of Escherichia coli as cloning vectors. Methods Enzymol. 1979;68:245–267. doi: 10.1016/0076-6879(79)68018-7. [DOI] [PubMed] [Google Scholar]
  2. Cantor C. R. Photo-cross-linking studies of nucleic acid structure. Ann N Y Acad Sci. 1980;346:379–385. doi: 10.1111/j.1749-6632.1980.tb22109.x. [DOI] [PubMed] [Google Scholar]
  3. Carlsson J., Drevin H., Axén R. Protein thiolation and reversible protein-protein conjugation. N-Succinimidyl 3-(2-pyridyldithio)propionate, a new heterobifunctional reagent. Biochem J. 1978 Sep 1;173(3):723–737. doi: 10.1042/bj1730723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cech T. R., Pardue M. L. Electron microscopy of DNA crosslinked with trimethylpsoralen: test of the secondary structure of eukaryotic inverted repeat sequences. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2644–2648. doi: 10.1073/pnas.73.8.2644. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chatterjee P. K., Cantor C. R. Photochemical production of psoralen - DNA monoadducts capable of subsequent photocrosslinking. Nucleic Acids Res. 1978 Oct;5(10):3619–3633. doi: 10.1093/nar/5.10.3619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cole R. S. Light-induced cross-linking of DNA in the presence of a furocoumarin (psoralen). Studies with phage lambda, Escherichia coli, and mouse leukemia cells. Biochim Biophys Acta. 1970 Sep 17;217(1):30–39. doi: 10.1016/0005-2787(70)90119-x. [DOI] [PubMed] [Google Scholar]
  7. Dale R. M., Ward D. C. Mercurated polynucleotides: new probes for hybridization and selective polymer fractionation. Biochemistry. 1975 Jun 3;14(11):2458–2469. doi: 10.1021/bi00682a028. [DOI] [PubMed] [Google Scholar]
  8. Dall'Acqua F., Marciani S., Rodighiero G. Inter-strand cross-linkages occurring in the photoreaction between psoralen and DNA. FEBS Lett. 1970 Jul 29;9(2):121–123. doi: 10.1016/0014-5793(70)80330-1. [DOI] [PubMed] [Google Scholar]
  9. Grassetti D. R., Murray J. F., Jr Determination of sulfhydryl groups with 2,2'- or 4,4'-dithiodipyridine. Arch Biochem Biophys. 1967 Mar;119(1):41–49. doi: 10.1016/0003-9861(67)90426-2. [DOI] [PubMed] [Google Scholar]
  10. Hayward G. S., Smith M. G. The chromosome of bacteriophage T5. I. Analysis of the single-stranded DNA fragments by agarose gel electrophoresis. J Mol Biol. 1972 Feb 14;63(3):383–395. doi: 10.1016/0022-2836(72)90435-4. [DOI] [PubMed] [Google Scholar]
  11. Isaacs S. T., Shen C. K., Hearst J. E., Rapoport H. Synthesis and characterization of new psoralen derivatives with superior photoreactivity with DNA and RNA. Biochemistry. 1977 Mar 22;16(6):1058–1064. doi: 10.1021/bi00625a005. [DOI] [PubMed] [Google Scholar]
  12. Parker R. C., Watson R. M., Vinograd J. Mapping of closed circular DNAs by cleavage with restriction endonucleases and calibration by agarose gel electrophoresis. Proc Natl Acad Sci U S A. 1977 Mar;74(3):851–855. doi: 10.1073/pnas.74.3.851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Parsons B. J. Psoralen photochemistry. Photochem Photobiol. 1980 Dec;32(6):813–821. doi: 10.1111/j.1751-1097.1980.tb04061.x. [DOI] [PubMed] [Google Scholar]
  14. Song P. S., Tapley K. J., Jr Photochemistry and photobiology of psoralens. Photochem Photobiol. 1979 Jun;29(6):1177–1197. doi: 10.1111/j.1751-1097.1979.tb07838.x. [DOI] [PubMed] [Google Scholar]
  15. Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]
  16. Wilson G. A., Young F. E. Purification and properties of the BamHI endonuclease. Methods Enzymol. 1980;65(1):147–153. doi: 10.1016/s0076-6879(80)65020-4. [DOI] [PubMed] [Google Scholar]

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