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Journal of Bacteriology, May 2003, p. 3076-3080, Vol. 185, No. 10
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.10.3076-3080.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

The Escherichia coli Fis Protein Stimulates Bacteriophage {lambda} Integrative Recombination In Vitro

Dominic Esposito* and Gary F. Gerard{dagger}

Invitrogen Corporation, Frederick, Maryland 21704

Received 12 November 2002/ Accepted 2 March 2003

The Escherichia coli nucleoid-associated protein Fis was previously shown to be involved in bacteriophage lambda site-specific recombination in vivo, enhancing the levels of both integrative recombination and excisive recombination. While purified Fis protein was shown to stimulate in vitro excision, Fis appeared to have no effect on in vitro integration reactions even though a 15-fold drop in lysogenization frequency had previously been observed in fis mutants. We demonstrate here that E. coli Fis protein does stimulate integrative lambda recombination in vitro but only under specific conditions which likely mimic natural in vivo recombination more closely than the standard conditions used in vitro. In the presence of suboptimal concentrations of Int protein, Fis stimulates the rate of integrative recombination significantly. In addition, Fis enhances the recombination of substrates with nonstandard topologies which may be more relevant to the process of in vivo phage lambda recombination. These data support the hypothesis that Fis may play an essential role in lambda recombination in the host cell.

* Corresponding author. Mailing address: Protein Expression Laboratory, SAIC-Frederick, Inc., NCI-Frederick, Bldg. 325, P.O. Box B, Frederick, MD 21702. Phone: (301) 846-7376. Fax: (301) 846-6289. E-mail: domespo{at}ncifcrf.gov.

{dagger} Present address: Transgenomic, Inc., Germantown, Md.

Journal of Bacteriology, May 2003, p. 3076-3080, Vol. 185, No. 10
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.10.3076-3080.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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