which GFP is brighter?

Model Organism: Worms Methods & Reagents
1

Hi,

We are trying to make some GFP fusion genes. Previously, we used the GFP sequence from pPD95.79 and transformed worms by bombardment. The fluorescence is too weak to work with under Lumar fluorescent dissecting scope.
Has anyone compared the old GFP from fire’s kit with current EGFP, and bombardment with injection? How much can these two changes improve the brightness?

Thanks very much.

2

I think the strength of your promoter or the localization of your protein may be the problem. I’ve used the GFP from both pPD95.75 and pPD95.77 without a problem, both via injection and bombardment. If my protein was strong, I got very bright expression. If my promoter was weak the opposite was true. I am pretty sure that these GFP are codon-modified for C. elegans, so you may not get a brighter signal from a mammalian EGFP variant.