Engineering the luxCDABE genes from Photorhabdus luminescens to provide a bioluminescent reporter for constitutive and promoter probe plasmids and mini-Tn5 constructs.
The luxCDABE operon of Photorhabdus luminescens has been cloned and engineered as an easily mobilisable cassette flanked by sites for commonly used restriction enzymes. Constitutive and promoter probe plasmids utilising the P. luminescens luxCDABE have been constructed using a number of compatible replicons and antibiotic markers. Complementary to these plasmids, a range of promoterless and constitutive luxCDABE mini-Tn5 derivatives has been constructed. The potential of coupling mini-Tn5 luxCDABE promoter probe transposons with automated luminometry and photometry to screen for mutants that exhibit growth phase variation in gene expression is demonstrated.
Food Microbiology Section, School of Biological Sciences, University of Nottingham Leics, USA.
M K
MK
Winson
S
S
Swift
P J
PJ
Hill
C M
CM
Sims
G
G
Griesmayr
B W
BW
Bycroft
P
P
Williams
G S
GS
Stewart
D003001:Cloning, Molecular; D003227:Conjugation, Genetic; D004251:DNA Transposable Elements; D004755:Enterobacteriaceae / Q000235:genetics* / Q000254:growth & development; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter / Q000235:genetics*; D008163:Luminescent Measurements; D009876:Operon; D010783:Photometry; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D015398:Signal Transduction
D004251:DNA Transposable Elements