Reporter gene to test new promotor - (Feb/03/2011 )
I Discovered a new kind of promotor that i now want to test in Pichia pastoris. I want to design a construct of the promotor in front of the reporter gene. I use 10 kind of promotors and want to see differences between them. Which kind of reporter gene would you recommand which can be quantitatively measured and does not influence Pichia pastoris metabolism.
cheers and thanks in advance
The most quantitative one is probably luciferase. You need a luminometer to read it, but it is more quantitative than fluorescent proteins. There is a wide choice of luciferase constructs from several vendors including Promega and NEB. Some luciferase are then secreted, so you can think they will minimally influence Pichia metabolism, furthermore, you can read them on the medium and easily perform time-course experiments.
For more info, I suggest to follow the luciferase channel at my blog