DETECTION OF MESSENGER-RNA IN STREPTOMYCES CELLS BY WHOLE-CELL HYBRIDIZATION WITH DIGOXIGENIN-LABELED PROBES

被引：49

作者：

HAHN, D ^{[1
]}

AMANN, RI ^{[1
]}

ZEYER, J ^{[1
]}

机构：

[1] TECH UNIV MUNICH,LEHRSTUHL MIKROBIOL,W-8000 MUNICH 2,GERMANY

来源：

APPLIED AND ENVIRONMENTAL MICROBIOLOGY | 1993年 / 59卷 / 08期

关键词：

D O I：

10.1128/AEM.59.8.2753-2757.1993

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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.

引用

页码：2753 / 2757

页数：5

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