Cloning and expression of plasmid DNA sequences involved in Salmonella serotype typhimurium virulence

• Published in: Molecular microbiology Vol. 3; no. 6; p. 733
• Main Authors: Norel, F, Coynault, C, Miras, I, Hermant, D, Popoff, M Y
• Format: Journal Article
• Language: English
• Published: England 01-06-1989
• Subjects: Animals; Cloning, Molecular; Conjugation, Genetic; DNA Transposable Elements; DNA, Bacterial - genetics; Electrophoresis, Polyacrylamide Gel; Escherichia coli - genetics; Female; Gene Expression Regulation; Mice; Mice, Inbred Strains; Mutation; Plasmids; Restriction Mapping; Salmonella typhimurium - genetics; Salmonella typhimurium - pathogenicity; Serotyping; Transformation, Genetic; Virulence
• ISSN: 0950-382X
• DOI: 10.1111/j.1365-2958.1989.tb00222.x

A 22kb region of the 90kb virulence-associated plasmid, pIP1350, of Typhimurium strain C52 was cloned into the mobilizable vector pSUP202, yielding plasmid pIP1352. This recombinant plasmid restored full virulence to plasmidless strain C53 in a mouse model. Transposon Tn5 insertion mutagenesis demonstrated the existence of two DNA sequences in pIP1352 designated VirA and VirB, both of which are essential for the expression of virulence. A recombinant plasmid containing only the VirA and VirB regions markedly increased the virulence of the plasmidless strain C53, but did not confer full virulence. These results suggested that a third virulence-associated region might be present on pIP1352. Eleven proteins encoded by the 22kb insert sequence of pIP1352 were identified in Escherichia coli SE5000 maxicells. The VirA region encoded at least two proteins with apparent molecular weights of 71,000 and 28,000 and the VirB region encoded two proteins of 43,000 and 38,000.