Expression of firefly luciferase gene in Erwinia amylovora.
In this study we describe an efficient stable genetic transformation of the phytopathogenic bacterium Erwinia amylovora using a recombinant expression vector encoding the firefly luciferase gene of Photinus pyralis, which is further controlled by IPTG-inducible promoter. Stably transformed E. amylovora cells maintain the same infectivity as the wild-type strain and, after induction with IPTG, produce luciferase. Luminescence produced by the action of luciferase on an exogenous substrate was easily detectable by a simple and rapid bioluminescent assay (BL). The transformed E. amylovora strain maintains the same high emission level, even after passage in pears, until about 15 days post-infection. Our findings therefore show that the luciferase assay can be conveniently used to follow the bacterial movement in plant tissue and its dissemination in controlled environments.
Gentilomi
Giovanna
G
Department of Clinical and Experimental Medicine, University of Bologna, via Massarenti 9, 40138 Bologna, Italy.
Zeri
Lisa
L
Ghini
Severino
S
Zerbini
Marialuisa
M
Zuffi
Elisa
E
Musiani
Monica
M
Girotti
Stefano
S
D000818:Animals; D001517:Coleoptera; D044084:Erwinia amylovora / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors; D008156:Luciferases / Q000235:genetics*; D049449:Luminescence; D010783:Photometry; D031989:Pyrus / Q000382:microbiology; D014170:Transformation, Genetic
D008156:Luciferases