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APPLICATION OF SUBTRACTION HYBRIDIZATION FOR THE DEVELOPMENT OF A RHIZOBIUM-LEGUMINOSARUM BIOVAR PHASEOLI AND RHIZOBIUM-TROPICI GROUP-SPECIFIC DNA-PROBE

Biomedical Sciences Research Institute Computer Science Research Institute Environmental Sciences Research Institute Nanotechnology & Advanced Materials Research Institute

Streit, W, Bjourson, AJ, Cooper, JE and Werner, D (1993) APPLICATION OF SUBTRACTION HYBRIDIZATION FOR THE DEVELOPMENT OF A RHIZOBIUM-LEGUMINOSARUM BIOVAR PHASEOLI AND RHIZOBIUM-TROPICI GROUP-SPECIFIC DNA-PROBE. FEMS MICROBIOLOGY ECOLOGY, 13 (1). pp. 59-67. [Journal article]

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URL: http://www3.interscience.wiley.com/journal/119301428/abstract?CRETRY=1&SRETRY=0

Abstract

A combined subtraction hybridization and polymerase chain reaction/amplification technique was used to develop a DNA probe which was specific for the Rhizobium leguminosarum biovar phaseoli and the Rhizobium tropici group. Total genomic DNA preparations from Rhizobium leguminosarum biovar viciae, Rhizobium leguminosarum biovar trifolii, Rhizobium sp., Agrobacterium tumefaciens, Rhizobium fredii, Bradyrhizobium japonicum, Bradyrhizobium ssp. and Rhizobium meliloti were pooled and used as subtracter DNA against total genomic DNA from the Rhizobium leguminosarum biovar phaseoli strain KIM5s. Only one round of subtraction hybridization at 65 degrees C was necessary to remove all cross-hybridizing sequences. Dot blot hybridizations with total genomic DNA of the eight subtracter organisms and 29 bacteria of different groups confirmed the high specificity of the isolated DNA sequences. Dot blot hybridizations with total genomic DNA from ten different R. leguminosarum biovar phaseoli and R. tropici strains resulted in strong hybridization signals for all strains tested. The DNA probe for the R. tropici and R. leguminosarum biovar phaseoli group was used for dot blot hybridization with DNA extracts from three tropical and one boreal soil. When correlated with data from Most Probable Number analyses the probe was capable of detecting as low as 3X10(4) homologous indigenous rhizobia per g soil. The technique offers great benefits for the development of DNA probes for monitoring bacterial populations in environmental samples.

Item Type:Journal article
Faculties and Schools:Faculty of Life and Health Sciences
Faculty of Life and Health Sciences > School of Biomedical Sciences
Research Institutes and Groups:Biomedical Sciences Research Institute
Biomedical Sciences Research Institute > Stratified Medicine
ID Code:1952
Deposited By:Mrs Caroline Adams
Deposited On:30 Nov 2009 09:36
Last Modified:14 Oct 2013 15:52

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