Abstract
Detection of mRNA of the thiostrepton resistance gene ( tsr ) harbored by plasmid pIJ673 in Streptomyces violacelatus was achieved by whole-cell hybridization with digoxigenin-labeled in vitro transcripts followed by an antibody-alkaline phosphatase detection of the digoxigenin reporter molecule. Prior to hybridization, the cells had to be permeabilized by lysozyme, the detergent Nonidet P-40, and toluene. The permeability of the S. violacelatus cells for probes and the antibody-alkaline phosphatase conjugate was demonstrated by hybridization with digoxigenin-labeled, 16S rRNA-targeted oligonucleotides.
Keywords
DigoxigeninMolecular biologyBiologyAlkaline phosphataseMolecular probeThiostreptonGene expressionIn situ hybridizationBiochemistryGeneRNAEnzymeRibosome
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Publication Info
- Year
- 1993
- Type
- article
- Volume
- 59
- Issue
- 8
- Pages
- 2753-2757
- Citations
- 60
- Access
- Closed
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Cite This
Dittmar Hahn,
Rudolf Amann,
Josef Zeyer
(1993).
Detection of mRNA in <i>Streptomyces</i> Cells by Whole-Cell Hybridization with Digoxigenin-Labeled Probes.
Applied and Environmental Microbiology
, 59
(8)
, 2753-2757.
https://doi.org/10.1128/aem.59.8.2753-2757.1993
Identifiers
- DOI
- 10.1128/aem.59.8.2753-2757.1993