In situ reverse transcription-PCR for monitoring gene expression in individual Methanosarcina mazei S-6 cells.

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

In situ reverse transcription-PCR for monitoring gene expression in individual Methanosarcina mazei S-6 cells. / Lange, M; Tolker-Nielsen, Tim; Molin, S; Ahring, B K.

In: Applied and Environmental Microbiology, Vol. 66, No. 5, 2000, p. 1796-800.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Lange, M, Tolker-Nielsen, T, Molin, S & Ahring, BK 2000, 'In situ reverse transcription-PCR for monitoring gene expression in individual Methanosarcina mazei S-6 cells.', Applied and Environmental Microbiology, vol. 66, no. 5, pp. 1796-800.

APA

Lange, M., Tolker-Nielsen, T., Molin, S., & Ahring, B. K. (2000). In situ reverse transcription-PCR for monitoring gene expression in individual Methanosarcina mazei S-6 cells. Applied and Environmental Microbiology, 66(5), 1796-800.

Vancouver

Lange M, Tolker-Nielsen T, Molin S, Ahring BK. In situ reverse transcription-PCR for monitoring gene expression in individual Methanosarcina mazei S-6 cells. Applied and Environmental Microbiology. 2000;66(5):1796-800.

Author

Lange, M ; Tolker-Nielsen, Tim ; Molin, S ; Ahring, B K. / In situ reverse transcription-PCR for monitoring gene expression in individual Methanosarcina mazei S-6 cells. In: Applied and Environmental Microbiology. 2000 ; Vol. 66, No. 5. pp. 1796-800.

Bibtex

@article{e4a5add0bd4111dd8e02000ea68e967b,
title = "In situ reverse transcription-PCR for monitoring gene expression in individual Methanosarcina mazei S-6 cells.",
abstract = "An in situ reverse transcription-PCR protocol for detecting specific mRNA in Methanosarcina mazei S-6 is described. This method allowed us to detect heat shock-induced increases in the intracellular levels of the transcript of the universal stress gene dnaK. The cell walls of paraformaldehyde-fixed cells were permeabilized by a thermal cycling procedure or by lysozyme treatment, and the cellular DNA was removed with DNase. The cells were subjected to a seminested reverse transcription-PCR protocol in which a digoxigenin-labeled primer was used. Detection of the reporter molecule was based on the 2-hydroxy-3-naphtoic acid-2'-phenylanilide phosphate-Fast Red detection system and binding of anti-digoxigenin-alkaline phosphatase conjugate. Fluorescence in permeabilized cells increased after a heat shock compared to fluorescence in non-heat-shocked cells, and the increase corresponded to an increase in the level of the dnaK transcript.",
author = "M Lange and Tim Tolker-Nielsen and S Molin and Ahring, {B K}",
note = "Keywords: Cell Wall; DNA Primers; Digoxigenin; Escherichia coli Proteins; HSP70 Heat-Shock Proteins; Indicators and Reagents; Methanosarcina; Molecular Chaperones; Muramidase; RNA, Archaeal; RNA, Messenger; Reverse Transcriptase Polymerase Chain Reaction; Transcription, Genetic",
year = "2000",
language = "English",
volume = "66",
pages = "1796--800",
journal = "Applied and Environmental Microbiology",
issn = "0099-2240",
publisher = "American Society for Microbiology",
number = "5",

}

RIS

TY - JOUR

T1 - In situ reverse transcription-PCR for monitoring gene expression in individual Methanosarcina mazei S-6 cells.

AU - Lange, M

AU - Tolker-Nielsen, Tim

AU - Molin, S

AU - Ahring, B K

N1 - Keywords: Cell Wall; DNA Primers; Digoxigenin; Escherichia coli Proteins; HSP70 Heat-Shock Proteins; Indicators and Reagents; Methanosarcina; Molecular Chaperones; Muramidase; RNA, Archaeal; RNA, Messenger; Reverse Transcriptase Polymerase Chain Reaction; Transcription, Genetic

PY - 2000

Y1 - 2000

N2 - An in situ reverse transcription-PCR protocol for detecting specific mRNA in Methanosarcina mazei S-6 is described. This method allowed us to detect heat shock-induced increases in the intracellular levels of the transcript of the universal stress gene dnaK. The cell walls of paraformaldehyde-fixed cells were permeabilized by a thermal cycling procedure or by lysozyme treatment, and the cellular DNA was removed with DNase. The cells were subjected to a seminested reverse transcription-PCR protocol in which a digoxigenin-labeled primer was used. Detection of the reporter molecule was based on the 2-hydroxy-3-naphtoic acid-2'-phenylanilide phosphate-Fast Red detection system and binding of anti-digoxigenin-alkaline phosphatase conjugate. Fluorescence in permeabilized cells increased after a heat shock compared to fluorescence in non-heat-shocked cells, and the increase corresponded to an increase in the level of the dnaK transcript.

AB - An in situ reverse transcription-PCR protocol for detecting specific mRNA in Methanosarcina mazei S-6 is described. This method allowed us to detect heat shock-induced increases in the intracellular levels of the transcript of the universal stress gene dnaK. The cell walls of paraformaldehyde-fixed cells were permeabilized by a thermal cycling procedure or by lysozyme treatment, and the cellular DNA was removed with DNase. The cells were subjected to a seminested reverse transcription-PCR protocol in which a digoxigenin-labeled primer was used. Detection of the reporter molecule was based on the 2-hydroxy-3-naphtoic acid-2'-phenylanilide phosphate-Fast Red detection system and binding of anti-digoxigenin-alkaline phosphatase conjugate. Fluorescence in permeabilized cells increased after a heat shock compared to fluorescence in non-heat-shocked cells, and the increase corresponded to an increase in the level of the dnaK transcript.

M3 - Journal article

C2 - 10788341

VL - 66

SP - 1796

EP - 1800

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 5

ER -

ID: 8780459