The expression of the Photinus pyralis luciferase gene in Staphylococcus aureus Cowan I allows the development of a live amplifiable tool for immunodetection.
We expressed the luc gene, encoding luciferase from Photinus pyralis, in Staphylococcus aureus Cowan I downstream of the plasmid-borne promoter for protein A. Constitutive luciferase synthesis did not impair the growth rate of the host nor did it affect the stability of the plasmid. Light production started immediately after addition of luciferin. The kinetic profile is of the glowing rather than the peak type. Because S. aureus Cowan I produces large quantities of protein A, of which a substantial part becomes covalently attached to rigid cell walls, the bacterial cells could be specifically immobilized on a substrate to which immunoglobulin G molecules were adsorbed either directly or as secondary antibodies. Light production from these cells can be used as a reporter tool for the detection of antigen-antibody complexes. Fourfold amplifications of the emitted signals were obtained by in situ incubation of the bound cells in bacterial growth medium.
Steidler
L
L
Laboratory for Molecular Biology, Universiteit Gent, Belgium.
Yu
W
W
Fiers
W
W
Remaut
E
E
D000818:Animals; D001681:Biological Assay / Q000379:methods; D001517:Coleoptera / Q000201:enzymology* / Q000235:genetics*; D015870:Gene Expression; D005818:Genetic Engineering; D007118:Immunoassay / Q000379:methods; D007700:Kinetics; D008027:Light; D008156:Luciferases / Q000235:genetics*; D018462:Photobiology; D010957:Plasmids / Q000235:genetics; D013211:Staphylococcus aureus / Q000201:enzymology* / Q000235:genetics*; D014170:Transformation, Genetic
D008156:Luciferases