SOS for GST-fusion protein expression - (Aug/17/2007 )

Hi guys

I have 3 GST-fusion proteins and I cannot get good expression after IPTG induction. The GST by itself is expressed very well. I was wondering if anyone knows how to improve the GST-fusion proteins expression from E-coli? Adding glucose will help? Or should I change the bacterial host?

Many thnx

Hak

First of all, check that your problem is not due to bad codon bias in your sequence. Try this site: http://gcua.schoedl.de/
If the GST expresses well, then the problem isn't with the cells, the medium or the IPTG. I suspect you have low-frequency codons in the gene. The quickest fix is to transfect the plasmid into a strain like Rosetta.