pgex - (Jan/27/2009 )

HI,
HAS ANY ONE TRIED TO EXPRESS PGEX in the eucayotic cells or any recomendation?

saraarasus on Jan 27 2009, 02:49 AM said:

Yes, what is your specific question

The pGex vectors are not intended for expression in eukaryotic cells and does not contain an appropriate promoter. These vectors are designed for expression of your protein in bacteria for purification.

saraarasus on Jan 27 2009, 01:49 AM said:

Look up if your protein has rare codon (if mammalian), if so then Novagen's rosetta BL21 strain would be a good option. I have used BL21(DE3)pLysS (auto lysozyme production) and has given me fairly good result.

elution of basic proteins
I am using the PGEX system to purify a basic protein of PI 9. I am using a denaturing 2M urea based protocol that i came across in a paper. The protein seems to be binding the column but when I try to elute it with PBS having 10mM glutathione + 2M urea nothing comes off. Can someone suggest some variations I could try to recover it from the column. Will changing PH conditions help?

pras45 on Feb 5 2009, 03:42 AM said:

I think you'll find that Rosettas are prokaryotic...

So the basic answer to the original question is in posting from rkay.