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Infection and Immunity, February 2000, p. 986-989, Vol. 68, No. 2
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Use of Lambda Phage S and R Gene Products in an Inducible Lysis System for Vibrio cholerae- and Salmonella enterica Serovar Typhimurium-Based DNA Vaccine Delivery Systems

Vivek Jain and John J. Mekalanos*

Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115

Received 11 August 1999/Returned for modification 13 September 1999/Accepted 15 November 1999

Novel methods for adapting DNA vaccine technology to the prevention of mucosal diseases are greatly needed. Here we show that regulated expression of phage lambda lysis genes S and R causes dramatic lysis of both Vibrio cholerae and Salmonella enterica serovar Typhimurium cells with concomitant release of plasmid DNA into the surrounding media. We also used single and double DNase mutant strains to show that secreted V. cholerae DNases can adversely affect the integrity of DNA molecules released upon lysis. These results suggest that incorporation of lambda SR-mediated lysis constructs and DNA stabilizing mutations into candidate live attenuated bacterial vaccines offers a promising approach for the development of effective mucosal DNA delivery vectors for humans.

* Corresponding author. Mailing address: Department of Microbiology and Molecular Genetics, Harvard Medical School, Building D1-411, 200 Longwood Ave., Boston, MA 02115. Phone: (617) 432-1935. Fax: (617) 738-7664. E-mail: jmekalanos{at}hms.harvard.edu.

Infection and Immunity, February 2000, p. 986-989, Vol. 68, No. 2
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.




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