Monitoring analysis of VBNC state of Escherichia coli O157 in soil

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Many pathogenic or non pathogenic bacteria enter the "viable but nonculturable (VBNC) state". In recent years, a great amount of research has revealed what occurs in cells as they enter and exist in this state, physiologically and morphologically. An aim of this study is to identify genes involved in the VBNC state of Escherichia coli O157 by mainly proteome analysis. At the first time, the experimental condition to enter the VBNC state was constructed, that is the incubation in sterile water at 4℃. The culturable cells decreased from 10^7 cells/ml to 10^2 cells/ml during 56 days incubation, but viable cells were constant at 10^7 cells/ml. The protein profiles of cells between VBNC 0 day and 56 days incubation were compared using SDS-PAGE and two dimensional gel electrophoresis, but both profiles were not observed significant difference. From proteome analysis with LC/MS/MS, about 1000 genes were detected, but they could not be found out the relation to VBNC state. Using "KO collection", that is all genes knock-out mutants in E.coli, VBNC genes were tried to identify. For 480 out of 3800 mutant strains, their VBNC states were examined. The genes have not been detected yet.