1887

Abstract

is an intracellular pathogen capable of multiplying to high levels in macrophages. By protein analysis, only a few proteins have been shown previously to be expressed at high levels in macrophages relative to bacteria grown in culture media. To identify additional genes that show increased expression during intracellular growth, we developed a plasmid for use in based on the induction of expression of green fluorescent protein. Clones of subsp. were identified that were fluorescent only intracellularly and not when grown . Sequencing identified a range of genes comprising some such as that are already known to be expressed intracellularly and some novel targets. One of these newly identified regulated genes, FTN1472/FTT1564, was selected for further study. Isogenic mutants were generated in subsp. and subsp. by allelic replacement. Inactivation of the gene resulted in abolition of polyphosphate production by , strongly supporting the bioinformatic analysis, which had suggested that the gene may encode a polyphosphate kinase. The mutants exhibited defects for intracellular growth in macrophages and were attenuated in mice, indicating a key role for the putative polyphosphate kinase in the virulence of .

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2008-10-01
2019-10-19
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