Simultaneous detection of different Rhizobium strains marked with either the Escherichia coli gusA gene or the Pyrococcus furiosus celB gene.
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.
Sessitsch
A
A
Department of Microbiology, Wageningen Agricultural University, The Netherlands.
Wilson
K J
KJ
Akkermans
A D
AD
de Vos
W M
WM
D001105:Archaea / Q000201:enzymology / Q000235:genetics*; D002874:Chromosome Mapping; D004251:DNA Transposable Elements; D017753:Ecosystem; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005798:Genes, Bacterial; D005819:Genetic Markers; D005966:Glucuronidase / Q000235:genetics; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000235:genetics; D010944:Plants / Q000382:microbiology; D012231:Rhizobium / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D001616:beta-Galactosidase / Q000235:genetics
D004251:DNA Transposable Elements; D005819:Genetic Markers; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C083711:cellobiose phosphotransferase B; D001616:beta-Galactosidase; D005966:Glucuronidase