Can anyone help with protein expression in pGEX vectors?

Dennis J. Templeton djt2 at po.CWRU.Edu
Tue Oct 6 16:34:51 EST 1992


In a previous article, lhanson at moose.uvm.edu (lhanson) says:

>	I've been trying to express 2 cDNAs in pGEX-2T.  Both cDNAs are

>(Deleted sad but all too typical tale of poor expression or solubility of 
>(fusion proteins

We have had in one case a near-miraculous improvement in solubility by
changing the temperature of the induced culture.  At 37 degrees it was
about 5% soluble, at 30 degrees it was more than 50% soluble, and the
expression was the same.  25degrees was slightly better.  For this one
protein, our routine is now to grow uninduced to about O.D. 0.5, then
induce and shut off the heat, then shake overnight.  

In other cases of poor solubility, this trick has worked *a little*...

It seems worth a try...

good luck,

dennis



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