Detection of rhythmic bioluminescence from luciferase reporters in cyanobacteria.
The unicellular cyanobacterium Synechococcus elongatus PCC 7942 is the model organism for studying prokaryotic circadian rhythms. Although S. elongatus does not display an easily measurable overt circadian behavior, its gene expression is under circadian control; hence, a "behavior" is created by linking a cyanobacterial promoter to either the bacterial luxAB or firefly luc luciferase genes to create reporter fusions whose activity can be easily monitored by bioluminescence. Numerous vectors have been created in our lab for introducing luciferase reporter genes into the S. elongatus chromosome. A choice of methods and equipment to detect light production from the luciferase fusions provides a means for high-throughput, automated mutant screens as well as testing rhythms from two promoter fusions within the same cell culture.
Mackey
Shannon R
SR
Department of Biology, Texas A&M University, College Station, USA.
Ditty
Jayna L
JL
Clerico
Eugenia M
EM
Golden
Susan S
SS
D000818:Animals; D002940:Circadian Rhythm / Q000502:physiology*; D017930:Genes, Reporter; D005818:Genetic Engineering; D005822:Genetic Vectors; D049453:Luciferases, Bacterial / Q000235:genetics; D049409:Luciferases, Firefly / Q000235:genetics; D049449:Luminescence; D008163:Luminescent Measurements / Q000295:instrumentation / Q000379:methods; D010957:Plasmids / Q000235:genetics; D012588:Scintillation Counting / Q000295:instrumentation / Q000379:methods; D046940:Synechococcus / Q000235:genetics / Q000502:physiology*; D014170:Transformation, Genetic
D049409:Luciferases, Firefly; D049453:Luciferases, Bacterial