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Site-specific recombination in mammalian cells expressing the Int recombinase of bacteriophage HK022 – Site-specific recombination in mammalian cells promoted by a phage integrase

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Abstract

The int gene of bacteriophage HK022, coding for the integrase protein, was cloned in a mammalian expression vector downstream of the human cytomegalovirus (CMV) promoter. Green monkey kidney cells (COS-1) and mouse embryo fibroblast cells (NIH3T3) transiently transfected with the recombinant plasmid express the integrase protein. Co-transfection of this plasmid with reporter plasmids for site-specific recombination and PCR analyses show that the integrase promotes site-specific integration as well as excision. These reactions occurred without the need to supply integration host factor and excisionase, the accessory proteins that are required for integrase-promoted site-specific recombination in vitro as well as in the natural host Escherichia coli.

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References

  1. Abremski K & Gottesman S (1981) J. Mol. Biol. 153: 67–78

    PubMed  Google Scholar 

  2. Ausubel F, Bernt R, Kingston RE, Moore DD, Seidman JG, Smith JA & Struhl K (1987) Current Protocols in Molecular Biology. J. Wiley & Sons, Inc., New York

    Google Scholar 

  3. Craig NL (1988) Annu. Rev. Genet 22: 77–105

    PubMed  Google Scholar 

  4. Dorgai L, Sloan S & Weisberg RA (1998). J. Mol. Biol. 277: 1059–1070

    PubMed  Google Scholar 

  5. Dorgai L, Yagil E & Weisberg R (1995) J. Mol. Biol. 252: 178–188

    PubMed  Google Scholar 

  6. Gluzman Y (1981) Cell 23: 175–182

    PubMed  Google Scholar 

  7. Goodman SD, Nicholson SC & Nash HA (1992) Proc. Natl. Acad. Sci. USA 89: 11910–11914

    PubMed  Google Scholar 

  8. Hirt B (1967) J. Mol. Biol. 26: 365–369

    PubMed  Google Scholar 

  9. Jainchill JL, Aaronson SA & Todaro GJ (1969) J. Virol. 4: 549–553

    PubMed  Google Scholar 

  10. Kilby NJ, Snaith MR & Murray JAH (1993) TIG 9: 413–421

    PubMed  Google Scholar 

  11. Kolot M, Gottfried P & Yagil E (1996) Mol. Gen. Genet. 253: 362–369

    PubMed  Google Scholar 

  12. Kolot M & Yagil E (1994) Mol. Gen. Genet. 245: 623–627

    PubMed  Google Scholar 

  13. Landy A (1989) Annu. Rev. Biochem. 58: 913–949

    PubMed  Google Scholar 

  14. Landy A (1993) Curr. Opin. Genet. Dev. 3: p. 699–707

    PubMed  Google Scholar 

  15. Nagaraja R & Weisberg RA (1990) J. Bacteriol 172: 6540–6550

    PubMed  Google Scholar 

  16. Nash HA (1983) Methods Enzymol. 100: 210–216

    PubMed  Google Scholar 

  17. Pluck A (1996) Int. J. Exp. Pathol. 77: 269–278

    PubMed  Google Scholar 

  18. Rajewsky K, Gu H, Kuhn R, Betz UA, Muller W, Roes J & Schwenk F (1996) J. Clin. Invest. 98: S51–3

    Google Scholar 

  19. Rossant J & Nagy A (1995) Nat. Med. 1: 592–594

    PubMed  Google Scholar 

  20. Sadowski PD (1993) FASEB J. 7: 760–767

    PubMed  Google Scholar 

  21. Sambrook J, Fritsch EF & Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.

    Google Scholar 

  22. Sauer B (1993) Methods Enzymol. 225: 890–900

    PubMed  Google Scholar 

  23. Sauer B (1994) Curr. Opin. Biotechnol. 5: 521–527

    PubMed  Google Scholar 

  24. Sauer B & Henderson N (1988) Proc. Natl. Acad. Sci. USA 85: 5166–5170

    PubMed  Google Scholar 

  25. Sauer B & Westphal H (1996) In: Lonai P (ed.) (pp 373–382) Harwood Academic Publishers, Amsterdam

  26. Segall AM, Goodman SD & Nash HA (1994) EMBO J. 13: 4536–4548

    PubMed  Google Scholar 

  27. Yagil E, Dolev S, Oberto J, Kislev N, Ramaiah N & Weisberg RA (1989) J. Mol. Biol. 207: 695–717

    PubMed  Google Scholar 

  28. Yagil E, Dorgai L & Weisberg R (1995) J. Mol. Biol. 252: 163–177

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry, Tel-Aviv University, Tel-Aviv, 69978, Israel

    Mikhail Kolot, Nava Silberstein & Ezra Yagil

Authors
  1. Mikhail Kolot
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  2. Nava Silberstein
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  3. Ezra Yagil
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Kolot, M., Silberstein, N. & Yagil, E. Site-specific recombination in mammalian cells expressing the Int recombinase of bacteriophage HK022 – Site-specific recombination in mammalian cells promoted by a phage integrase. Mol Biol Rep 26, 207–213 (1999). https://doi.org/10.1023/A:1007096701720

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  • DOI: https://doi.org/10.1023/A:1007096701720

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