EGFP fusion protein expression - Expression of a GFP fusion protein in E.coli (Apr/17/2007 )
Hello,
Does someone know if it is possible to use a normal EGFP vector from clonetech to expres a GFP fusion protein in competent E.coli?
Or is this only possible in eukaryotic cells because of the wrong prokayrotic promotor?
If it is not possible, what kind of GFP vector do I have to use?
Greetz Roy
Does someone know if it is possible to use a normal EGFP vector from clonetech to expres a GFP fusion protein in competent E.coli?
Or is this only possible in eukaryotic cells because of the wrong prokayrotic promotor?
If it is not possible, what kind of GFP vector do I have to use?
Greetz Roy
I guess it should be possible, but the vectors for expressing N- or C- fusion proteins will have better multiple cloning sites downstream or upstream of the GFP.
Ceri
Do a search. Its been done.
I can't find the answar 
I think that it is not possible because of the HCMV promotor before the GFP gene.
There is a bacterial promotor, for transcribing the Kan. gene, but can you use this promotor also for transcribing GFP and the insert gene?
You can try the Gateway Cloning system. It lets you construct vectors with tags in your vector of interest. It also has vectors that express fusion tags.
Downside is..you'll have to purchase a lot of stuff from them. Or find some labs close to yours that use this system...borrowing works too!