Study on the conversion from VBNC to culturable state in Vibrio cholerae

Research Project

Project/Area Number	26460543
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Research Field	Bacteriology (including mycology)
Research Institution	National Center for Global Health and Medicine
Principal Investigator	Hamabata Takashi 国立研究開発法人国立国際医療研究センター, その他部局等, 細菌感染研究室長 (40311427)
Co-Investigator(Renkei-kenkyūsha)	Senoh Mitsutoshi 国立感染症研究所, 細菌第二部, 主任研究官 (20646624)
Project Period (FY)	2014-04-01 – 2017-03-31
Project Status	Completed (Fiscal Year 2016)
Budget Amount *help	¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000) Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000) Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000) Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywords	コレラ菌 / カタラーゼ / VBNC / RNAマイクロアレイ / 培養可能状態
Outline of Final Research Achievements	The viable but nonculturable (VBNC) Vibrio cholerae converts to culturable state by co-culture of with cultured cells. The converting factor (FCVC) was purified from HT-29 cells through several kinds of column chromatography. The final purified single band on SDS-PAGE was analyzed by nano-LC MS/MS and turned out to be human catalase. Both catalase RNAi analysis of HT-29 cells and catalase inhibitor 3-amino-1,2,4-triazole greatly reduced the converting activity of crude cell extract, confirming that FCVC is catalase. To identify the genes that involve in the conversion, VBNC V. cholerae was collected by centrifugation, treated with catalase and bacterial RNA was purified on several different time points. Since RNA microarray analysis revealed too many genes responded to catalase, negative control RNA should be compared. However, low conversion was always seen even without catalase, which reason turned out to be the condensation of bacteria by centrifugation.