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Science 26 September 1997:
Vol. 277. no. 5334, p. 1901
DOI: 10.1126/science.277.5334.1901r
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Many major bacterial pathogens live within the cells of the host to avoid some of the host's immune protections. If the bacterial genes that are vital for their survival were known more options would exist for designing potential antimicrobial therapies. Valdivia and Falkow have developed a method to select for such genes. They constructed a selection plasmid that contains the fluorescent green protein gene and pieces of the Salmonella typhimurium genome. After transfection into S. typhimurium, the bacteria were allowed to invade macrophages. Bacteria from fluorescent macrophages were grown in agar and then only those that did not fluoresce were used to reinfect mammalian cells. This simple system selects for genes preferentially expressed while the bacterium is intracellular and has the potential to be used with bacteria that have as yet intractable genetic systems.

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