PMID: 9546162Apr 18, 1998Paper

Characterization of the rrnB operon of the plant pathogen Rhodococcus fascians and targeted integrations of exogenous genes at rrn loci

Applied and Environmental Microbiology
A PisabarroJ F Martín

Abstract

A 6.0-kb SalI DNA fragment containing an entire rRNA operon (rrnB) was cloned from a cosmid gene bank of the phytopathogenic strain Rhodococcus fascians D188. The nucleotide sequence of the 6-kb fragment was determined and had the organization 16S rRNA-spacer-23S rRNA-spacer-5S rRNA without tRNA-encoding genes in the spacer regions. The 5' and 3' ends of the mature 16S, 23S, and 5S rRNAs were determined by alignment with the rrn operons of Bacillus subtilis and other gram-positive bacteria. Four copies of the rrn operons were identified by hybridization with an rrnB probe in R. fascians type strain ATCC 12974 and in the virulent strain R. fascians D188. However, another isolate, CECT 3001 (= NRRL B15096), also classified as R. fascians, produced five rrn-hybridizing bands. An integrative vector containing a 2.5-kb DNA fragment internal to rrnB was constructed for targeted integration of exogenous genes at the rrn loci. Transformants carrying the exogenous chloramphenicol resistance gene (cmr) integrated in different rrn operons were obtained. These transformants had normal growth rates in complex medium and minimal medium and were fully stable for the integrated marker.

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Citations

Sep 29, 2004·FEMS Microbiology Reviews·Volker GürtlerRobert Seviour
Mar 13, 2001·DNA and Cell Biology·K C HoT L Chung
Jun 19, 2010·FEMS Microbiology Ecology·Allison E RayTimothy S Magnuson
Sep 29, 2001·Journal of Applied Microbiology·R D Bagsic-OpulenciaM Fegan
Feb 14, 2004·Letters in Applied Microbiology·M CandelaP Brigidi
Nov 10, 2001·Annual Review of Phytopathology·K GoethalsM Holsters
Jun 3, 1999·Journal of Microbiological Methods·J García-MartínezF Rodríguez-Valera
Apr 1, 2000·Applied and Environmental Microbiology·J A KlappenbachT M Schmidt

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