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J Bacteriol, April 1998, p. 2063-2071, Vol. 180, No. 8
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Use of Bacteriophage lambda  Recombination Functions To Promote Gene Replacement in Escherichia coli

Kenan C. Murphy*

Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, Worcester, Massachusetts 01655

Received 1 October 1997/Accepted 6 February 1998

Replacement of Escherichia coli's RecBCD function with phage lambda 's Red function generates a strain whose chromosome recombines with short linear DNA fragments at a greatly elevated rate. The rate is at least 70-fold higher than that exhibited by a recBC sbcBC or recD strain. The value of the system is highlighted by gene replacement with a PCR-generated DNA fragment. The Delta recBCD::Plac-red kan replacement allele can be P1 transduced to other E. coli strains, making the hyper-Rec phenotype easily transferable.

* Mailing address: Molecular Genetics and Microbiology, University of Massachusetts Medical Center, Worcester, MA 01655. Phone: (508) 856-6069. Fax: (508) 856-5920. E-mail: kenan.murphy{at}ummed.edu.




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