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Applied and Environmental Microbiology, August 2001, p. 3735-3738, Vol. 67, No. 8
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.8.3735-3738.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Use of an Intergenic Region in Pseudomonas syringae pv. Syringae B728a for Site-Directed Genomic Marking of Bacterial Strains for Field Experiments

Susan S. Hirano,1,* David K. Willis,1,2 Murray K. Clayton,1,3 and Christen D. Upper1,2

Departments of Plant Pathology1 and Statistics,3 University of Wisconsin---Madison, and Plant Disease Resistance Research Unit, Agricultural Research Service, U.S. Department of Agriculture,2 Madison, Wisconsin 53706

Received 8 January 2001/Accepted 30 May 2001

To construct differentially-marked derivatives of our model wild-type strain, Pseudomonas syringae pv. syringae B728a (a causal agent of bacterial brown spot disease in snap bean plants), for field experiments, we selected a site in the gacS-cysM intergenic region for site-directed insertion of antibiotic resistance marker cassettes. In each of three field experiments, population sizes of the site-directed chromosomally marked B728a derivatives in association with snap bean plants were not significantly different from that of the wild-type strain. Inserts of up to 7 kb of DNA in the intergenic region did not measurably affect fitness of B728a in the field. The site is useful for site-directed genomic insertions of single copies of genes of interest.

* Corresponding author. Mailing address: Department of Plant Pathology, University of Wisconsin---Madison, 1630 Linden Drive, Madison, WI 53706. Phone: (608) 262-7236. Fax: (608) 263-2626. E-mail: ssh{at}plantpath.wisc.edu.

Applied and Environmental Microbiology, August 2001, p. 3735-3738, Vol. 67, No. 8
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.8.3735-3738.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

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