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Microbiology 144 (1998), 3205-3218; DOI  10.1099/00221287-144-11-3205
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Isolation of carbon- and nitrogen-deprivation-induced loci of Sinorhizobium meliloti 1021 by Tn5-luxAB mutagenesis

Anne Milcamps1,2, Daniel M. Ragatz1,2, PyungOk Lim1,2, Kelly A. Berger2 and Frans J. de Bruijin1,2,3,*

NSF Center for Microbial Ecology, University, East Lansing, MI 48824, USA
MSU-DOE Plant Research Laboratory, Rm 306, Plant Biology Building University, East Lansing, MI 48824, USA
Department of Microbiology, Michigan State University, East Lansing, MI 48824, USA

*Author for correspondence: Frans J. de Bruijn. Tel: + 1 517 353 2229. Fax: + 1 517 355 9614.e-mail: debruijn@pilot.msu.edu

ABSTRACT

SUMMARY: Soil bacteria, such as Sinorhizobium meliloti, are subject to variation in environmental conditions, including carbon- and nitrogen-deprivation. The ability of bacteria to sense changes in their environment and respond accordingly is of vital importance to their survival and persistence in the soil and rhizosphere. A derivative of Tn5 which creates transcriptional fusions to the promoterless luxAB genes was used to mutagenize 5. meliloti 1021 and 5000 insertion mutants were subsequently screened for gene fusions induced by selected environmental stresses. The isolation of 21 gene fusions inducedby nitrogen-deprivation and 12 induced by carbon-deprivation is described. Cloning and partial DNA sequence analysis of the transposon-tagged loci revealed a variety of novel genes, as well as S. meliloti genes with significant similarity to known bacterial loci. In addition, nodule occupancy studies were carried out with selected TnSluxAB insertion mutants to examine the role of the tagged genes in competition.

Keywords: Rhizobium starvation, nutrient-deprivation, luciferase activity, Tn5-lux, competition




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