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. 1992 Sep 11;20(17):4451–4455. doi: 10.1093/nar/20.17.4451

Stoichiometry of the Cre recombinase bound to the lox recombining site.

A Mack 1, B Sauer 1, K Abremski 1, R Hoess 1
PMCID: PMC334171  PMID: 1408747

Abstract

The site-specific recombinase Cre from bacteriophage P1 binds and carries out recombination at a 34 bp lox site. The lox site consists of two 13 bp inverted repeats, separated by an 8 bp spacer region. Both the palindromic nature of the site and the results of footprinting and band shift experiments suggest that a minimum of two Cre molecules bind to a lox site. We report here experiments that demonstrate the absolute stoichiometry of the Cre-lox complex to be one molecule of Cre bound per inverted repeat, or two molecules per lox site.

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Selected References

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  1. Abremski K., Hoess R. Bacteriophage P1 site-specific recombination. Purification and properties of the Cre recombinase protein. J Biol Chem. 1984 Feb 10;259(3):1509–1514. [PubMed] [Google Scholar]
  2. Craig N. L. The mechanism of conservative site-specific recombination. Annu Rev Genet. 1988;22:77–105. doi: 10.1146/annurev.ge.22.120188.000453. [DOI] [PubMed] [Google Scholar]
  3. Granger-Schnarr M., Lloubes R., de Murcia G., Schnarr M. Specific protein-DNA complexes: immunodetection of the protein component after gel electrophoresis and Western blotting. Anal Biochem. 1988 Oct;174(1):235–238. doi: 10.1016/0003-2697(88)90540-4. [DOI] [PubMed] [Google Scholar]
  4. Hendrickson W., Schleif R. F. Regulation of the Escherichia coli L-arabinose operon studied by gel electrophoresis DNA binding assay. J Mol Biol. 1984 Sep 25;178(3):611–628. doi: 10.1016/0022-2836(84)90241-9. [DOI] [PubMed] [Google Scholar]
  5. Hoess R. H., Abremski K. Interaction of the bacteriophage P1 recombinase Cre with the recombining site loxP. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1026–1029. doi: 10.1073/pnas.81.4.1026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hoess R., Abremski K., Irwin S., Kendall M., Mack A. DNA specificity of the Cre recombinase resides in the 25 kDa carboxyl domain of the protein. J Mol Biol. 1990 Dec 20;216(4):873–882. doi: 10.1016/S0022-2836(99)80007-2. [DOI] [PubMed] [Google Scholar]
  7. Hope I. A., Struhl K. GCN4, a eukaryotic transcriptional activator protein, binds as a dimer to target DNA. EMBO J. 1987 Sep;6(9):2781–2784. doi: 10.1002/j.1460-2075.1987.tb02573.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  9. Sauer B., Henderson N. Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1. Proc Natl Acad Sci U S A. 1988 Jul;85(14):5166–5170. doi: 10.1073/pnas.85.14.5166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  11. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Wierzbicki A., Kendall M., Abremski K., Hoess R. A mutational analysis of the bacteriophage P1 recombinase Cre. J Mol Biol. 1987 Jun 20;195(4):785–794. doi: 10.1016/0022-2836(87)90484-0. [DOI] [PubMed] [Google Scholar]
  13. Yang C. C., Nash H. A. The interaction of E. coli IHF protein with its specific binding sites. Cell. 1989 Jun 2;57(5):869–880. doi: 10.1016/0092-8674(89)90801-5. [DOI] [PubMed] [Google Scholar]

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