Differential fluorescence induction analysis of Streptococcus pneumoniae identifies genes involved in pathogenesis

被引:77
作者
Marra, A [1 ]
Asundi, J [1 ]
Bartilson, M [1 ]
Lawson, S [1 ]
Fang, F [1 ]
Christine, J [1 ]
Wiesner, C [1 ]
Brigham, D [1 ]
Schneider, WP [1 ]
Hromockyj, AE [1 ]
机构
[1] Prot Design Labs Inc, Fremont, CA 94555 USA
关键词
D O I
10.1128/IAI.70.3.1422-1433.2002
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Differential fluorescence induction (DFI) technology was used to identify promoters of Streptococcus pneumoniae induced under various in vitro and in vivo conditions. A promoter-trap library using green fluorescent protein as the reporter was constructed in S. pneumoniae, and the entire library was screened for clones exhibiting increased gfp expression under the chosen conditions. The in vitro conditions used were chosen to mimic aspects of the in vivo environment encountered by the pathogen once it enters a host: changes in temperature, osmolarity, oxygen, and iron concentration, as well as blood. In addition, the library was used to infect animals in three different models, and clones induced in these environments were identified. Several promoters were identified in multiple screens, and genes whose promoters were induced twofold or greater under the inducing condition were mutated to assess their roles in virulence. A total of 25 genes were mutated, and the effects of the mutations were assessed in at least two different infection models. Over 50% of these mutants were attenuated in at least one infection model. We show that DFI is a useful tool for identifying bacterial virulence factors as well as a means of elucidating the microenvironment encountered by pathogens upon infection.
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收藏
页码:1422 / 1433
页数:12
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