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.
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.
Jain
V
V
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.
Mekalanos
J J
JJ
D001428:Bacterial Vaccines / Q000008:administration & dosage*; D010582:Bacteriophage lambda / Q000235:genetics*; D022121:Cholera Vaccines / Q000008:administration & dosage*; D004269:DNA, Bacterial / Q000378:metabolism; D003851:Deoxyribonucleases / Q000494:pharmacology; D010957:Plasmids; D012486:Salmonella typhimurium / Q000276:immunology*; D019444:Vaccines, DNA / Q000008:administration & dosage*
D001428:Bacterial Vaccines; D022121:Cholera Vaccines; D004269:DNA, Bacterial; D019444:Vaccines, DNA; D003851:Deoxyribonucleases