Staphylococcus aureus cell extract transcription-translation assay: firefly luciferase reporter system for evaluating protein translation inhibitors.
The promoter for the Staphylococcus aureus capsule polysaccharide synthesis gene (cap1A) was cloned in front of the firefly luciferase gene for use in a cell extract S. aureus transcription-translation system. The assay is rapid, reproducible, and sensitive and has a lower background level than the radiolabeled amino acid incorporation translation assays. We present data evaluating a transcription inhibitor and a number of protein translation inhibitors in this system.
Murray
R W
RW
Pharmacia Corporation, Kalamazoo, Michigan 49007, USA.
Melchior
E P
EP
Hagadorn
J C
JC
Marotti
K R
KR
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D003001:Cloning, Molecular; D004353:Drug Evaluation, Preclinical / Q000379:methods; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics*; D008826:Microbial Sensitivity Tests; D010957:Plasmids; D014176:Protein Biosynthesis / Q000187:drug effects*; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects*
D000900:Anti-Bacterial Agents; D008156:Luciferases