Construction and evaluation of a eukaryotic expression plasmid for stable delivery using attenuated Salmonella


Garmory, HS; Titball, RW; Brown, KA; Bennett, AM; (2003) Construction and evaluation of a eukaryotic expression plasmid for stable delivery using attenuated Salmonella. Microbial pathogenesis, 34 (3). pp. 115-119. ISSN 0882-4010 DOI: 10.1016/s0882-4010(02)00176-6

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Abstract

An approach to enhancing the stability of eukaryotic expression plasmids for delivery using attenuated Salmonella has been evaluated. The expression apparatus and beta-galactosidase gene from the expression plasmid, pCMVbeta, was cloned into the low copy number plasmid pLG339. The resulting construct, pLGbetaGAL, was shown to have a lower copy number than pCMVbeta in Salmonella enterica var Typhimurium aroA strain SL7207. Furthermore, beta-galactosidase-specific antibody was induced in mice following intramuscular inoculation with pLGbetaGAL as naked DNA. Following oral administration of mice with SL7207/pCMVbeta, recombinants could not be detected in tissues 3 days after inoculation. In comparison, SL7207/pLGbetaGAL recombinant bacteria could be detected in the Peyer's patches and spleens indicating that the Salmonella strain was stable. However, both SL7207/pCMVbeta and SL7207/pLGbetaGAL failed to induce beta-galactosidase-specific IgG in vivo. The mechanism by which attenuated Salmonella are able to release heterologous DNA for antigen processing and presentation is not yet understood. These results suggest that the mechanism needs to be further elucidated in order to rationally improve the system. (C) 2003 Elsevier Science Ltd. All rights reserved.

Item Type: Article
Keywords: Salmonella, expression plasmid, copy number, Typhimurium, vaccination, vaccines, systems, vectors, cells, Animal, Eukaryotic Cells, Female, Gene Dosage, Genetic Vectors, Human, Immunization, Immunoglobulin G, blood, Mice, Mice, Inbred BALB C, Plasmids, Salmonella typhimurium, genetics, metabolism, pathogenicity, Vaccines, Attenuated, Vaccines, DNA, beta-Galactosidase, genetics, immunology, metabolism
Faculty and Department: Faculty of Infectious and Tropical Diseases > Dept of Pathogen Molecular Biology
PubMed ID: 12631472
Web of Science ID: 181884400001
URI: http://researchonline.lshtm.ac.uk/id/eprint/17740

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