Detection of mRNA in Streptomyces Cells by Whole-Cell Hybridization with Digoxigenin-Labeled Probes-Reference-Cited by-同舟云学术

Detection of mRNA in Streptomyces Cells by Whole-Cell Hybridization with Digoxigenin-Labeled Probes

Published:1993-08 Issue:8 Volume:59 Page:2753-2757
ISSN:0099-2240
Container-title:Applied and Environmental Microbiology
language:en
Short-container-title:Appl Environ Microbiol

Author:

Hahn Dittmar¹,Amann Rudolf I.¹,Zeyer Josef¹

Affiliation:

1. Institute of Terrestrial Ecology, Soil Biology, Eidgenössische Technische Hochschule Zürich, Grabenstrasse 3, CH-8952 Schlieren, Switzerland, and Lehrstuhl für Mikrobiologie, Technische Universität München, Arcisstrasse 21, D-8000 Munich 2, Germany2

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.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Link

https://journals.asm.org/doi/pdf/10.1128/aem.59.8.2753-2757.1993

Reference32 articles.

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