Reconstructing promoter activity from Lux bioluminescent reporters.

摘要: The bacterial Lux system is used as a gene expression reporter. It fast, sensitive and non-destructive, enabling high frequency measurements. Originally developed for cells, it has also been adapted eukaryotic can be whole cell biosensors, or in real time with live animals without the need euthanasia. However, correct interpretation of bioluminescent data limited: bioluminescence different from because nonlinear molecular enzyme dynamics system. We have computational approach that, first time, allows users assays to infer transcription levels light output. This based upon new mathematical model activity, that includes actions LuxAB, LuxEC Fre, improved mechanisms all reactions, well synthesis turn-over proteins. calibrated experimental LuxAB Fre reactions Photorhabdus luminescens-the source modern reporters-while literature LuxEC. Importantly, show clear evidence previously unreported product inhibition reaction. Model simulations predicted profiles very changes expression, transient peaks output, similar output seen some sets. By incorporating into Bayesian inference scheme, we reverse engineer promoter activity bioluminescence. examples where decrease would better interpreted switching off promoter, an increase longer period expression. could benefit technology.