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Journal of Bacteriology, March 1999, p. 1733-1738, Vol. 181, No. 6
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Physiological States of Individual Salmonella typhimurium Cells Monitored by In Situ Reverse Transcription-PCR

Kim Holmstrøm,1 Tim Tolker-Nielsen,2 and Søren Molin2,*

Biotechnological Institute, DK-2970 Hørsholm,1 and Department of Microbiology, The Technical University of Denmark, DK-2800 Lyngby,2 Denmark

Received 8 July 1998/Accepted 25 November 1998

The possibility of using levels of specific mRNAs in individual bacteria as indicators of single-cell physiology was investigated. Estimates of the numbers of groEL and tsf mRNAs per cell in Salmonella typhimurium cells in different physiological states were obtained by Northern analysis. The average number of groEL mRNAs per cell was estimated to be 22 in fast-growing cultures and 197 in heat-shocked cultures. The average number of tsf mRNAs per cell was estimated to be 37 in fast-growing cultures, 4 in slow-growing cultures, and 0 in nongrowing cultures. The potential of mRNA-targeted in situ reverse transcription (RT)-PCR to monitor quantitatively different levels of groEL and tsf mRNA in individual cells and thus monitor both specific gene induction and general growth activity was assessed. Neither groEL nor tsf mRNA was present in stationary-phase cells, but it was shown that stationary-phase cells contain other RNA species at high levels, which may provide a possibility for monitoring directly stationary-phase individual cells by the use of in situ RT-PCR. The outcome of the in situ RT-PCR analyses indicated that a population of fast-growing cells is heterogeneous with respect to groEL mRNA single-cell contents, suggesting a cell-cycle-controlled expression of groEL in S. typhimurium, whereas a fast-growing culture is homogeneous with respect to tsf mRNA single-cell contents, suggesting that the level of tsf mRNA is relatively constant during the cell cycle.

* Corresponding author. Mailing address: Department of Microbiology, Bldg. 301, The Technical University of Denmark, DK-2800 Lyngby, Denmark. Phone: 45-45252513. Fax: 45-45887328. E-mail: sm{at}im.dtu.dk.

Journal of Bacteriology, March 1999, p. 1733-1738, Vol. 181, No. 6
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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