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Chapter 16 : Regulation of Virulence Gene Expression in Vivo

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Abstract:

In this chapter, the invasion locus of serovar Typhimurium is used to exemplify several general points about the regulation of virulence gene expression in vivo. The chapter also includes a brief description of in vivo expression technology (IVET), designed to select for bacterial genes that are transcriptionally induced in the host. The regulation of invasion gene expression involves an increasingly large number of regulatory loci in addition to HilA and InvF. The original IVET system was based on the fact that mutants of serovar Typhimurium are completely incapable of surviving within the animal host. PurA is required at all stages of infection in all host tissues. Researchers designed an analogous system using the chloramphenicol acetyl-transferase or cat gene as a reporter in place of . A number of researchers have subsequently developed IVET systems that are variations on a theme and have used these systems to identify in vivo-induced genes in a variety of both prokaryotic and eukaryotic pathogens. These IVET systems fall into three categories: (i) selection systems based on metabolic (e.g., ) or antibiotic (e.g., cat) reporters, (ii) recombination-based systems, and (iii) green fluourescent protein (GFP)-based systems. Approximately 10% of the genes that answered these selections encode products that are known or thought to be involved in serovar Typhimurium pathogenesis.

Citation: Slauch J. 2000. Regulation of Virulence Gene Expression in Vivo, p 241-249. In Brogden K, Roth J, Stanton T, Bolin C, Minion F, Wannemuehler M (ed), Virulence Mechanisms of Bacterial Pathogens, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818111.ch16

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Type III Secretion System
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Gene Expression and Regulation
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Figures

Image of FIGURE 1
FIGURE 1

Model for the regulation of the serovar Typhimurium invasion apparatus. The boxed genes are located in SPI1. Arrows from regulatory genes indicate activation, whereas lines ending in short lines indicate repression. It is not clear in most cases whether the action is direct or indirect.

Citation: Slauch J. 2000. Regulation of Virulence Gene Expression in Vivo, p 241-249. In Brogden K, Roth J, Stanton T, Bolin C, Minion F, Wannemuehler M (ed), Virulence Mechanisms of Bacterial Pathogens, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818111.ch16
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Image of FIGURE 2
FIGURE 2

Construction of pIVET1 fusions. Random chromosomal fragments were cloned 5′ to the promoterless gene in pIVET1. The plasmid is based on oriR6K. Therefore, replication is dependent on the Pi protein, which must be supplied in trans. When the plasmids are introduced into a Pi strain of serovar Typhimurium, they must integrate into the chromosome by homologous recombination with the cloned fragment to be stably maintained. This generates a tandem duplication where one promoter drives the fusion, while the other promoter drives the wild-type copy of the gene of interest.

Citation: Slauch J. 2000. Regulation of Virulence Gene Expression in Vivo, p 241-249. In Brogden K, Roth J, Stanton T, Bolin C, Minion F, Wannemuehler M (ed), Virulence Mechanisms of Bacterial Pathogens, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818111.ch16
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Image of FIGURE 3
FIGURE 3

Screen for tissue-specific gene expression using IVET fusions. The figure shows the predicted outcome for a fusion that is specifically induced in the small intestine. The in vivo induced fusion strain is phenotypically Lac (open circles) on lactose MacConkey agar. The constitutive fusion strain is Lac (solid circles).

Citation: Slauch J. 2000. Regulation of Virulence Gene Expression in Vivo, p 241-249. In Brogden K, Roth J, Stanton T, Bolin C, Minion F, Wannemuehler M (ed), Virulence Mechanisms of Bacterial Pathogens, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818111.ch16
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References

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Tables

Generic image for table
TABLE 1

IVET systems and selections

Citation: Slauch J. 2000. Regulation of Virulence Gene Expression in Vivo, p 241-249. In Brogden K, Roth J, Stanton T, Bolin C, Minion F, Wannemuehler M (ed), Virulence Mechanisms of Bacterial Pathogens, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818111.ch16
Generic image for table
TABLE 2

Competition between in vivo-induced IVET fusions and a constitutive IVET fusion

Citation: Slauch J. 2000. Regulation of Virulence Gene Expression in Vivo, p 241-249. In Brogden K, Roth J, Stanton T, Bolin C, Minion F, Wannemuehler M (ed), Virulence Mechanisms of Bacterial Pathogens, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818111.ch16

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