Abstract
A new marker system for gram-negative bacteria was developed on the basis of the celB gene from the hyperthermophilic archaeon Pyrococcus furiosus, which encodes a thermostable beta-glucosidase with a high level of beta-galactosidase activity. The celB gene is highly suitable as a marker for studying plant-bacterium interaction because endogenous background beta-glucosidase and beta-galactosidase enzyme activity can readily be inactivated by heat and because inexpensive substrates for detection are commercially available. Two celB-expressing transposons were constructed for use in ecological studies of a variety of gram-negative bacteria. The combined use of the gusA marker gene and celB allowed the simultaneous detection of several Rhizobium strains on a plant, and multiple-strain occupancy of individual modules also could be easily detected.
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Selected References
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- De Leij F., Sutton E. J., Whipps J. M., Fenlon J. S., Lynch J. M. Impact of Field Release of Genetically Modified Pseudomonas fluorescens on Indigenous Microbial Populations of Wheat. Appl Environ Microbiol. 1995 Sep;61(9):3443–3453. doi: 10.1128/aem.61.9.3443-3453.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hattemer-Frey H. A., Brandt E. J., Travis C. C. Small-scale field test of the genetically engineered lacZY marker. Regul Toxicol Pharmacol. 1990 Jun;11(3):253–261. doi: 10.1016/0273-2300(90)90025-7. [DOI] [PubMed] [Google Scholar]
- Voorhorst W. G., Eggen R. I., Luesink E. J., de Vos W. M. Characterization of the celB gene coding for beta-glucosidase from the hyperthermophilic archaeon Pyrococcus furiosus and its expression and site-directed mutation in Escherichia coli. J Bacteriol. 1995 Dec;177(24):7105–7111. doi: 10.1128/jb.177.24.7105-7111.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilson K. J., Sessitsch A., Corbo J. C., Giller K. E., Akkermans A. D., Jefferson R. A. beta-Glucuronidase (GUS) transposons for ecological and genetic studies of rhizobia and other gram-negative bacteria. Microbiology. 1995 Jul;141(Pt 7):1691–1705. doi: 10.1099/13500872-141-7-1691. [DOI] [PubMed] [Google Scholar]
- de Lorenzo V., Herrero M., Jakubzik U., Timmis K. N. Mini-Tn5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in gram-negative eubacteria. J Bacteriol. 1990 Nov;172(11):6568–6572. doi: 10.1128/jb.172.11.6568-6572.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]