[ccp4bb]: problem with SeMet expression

[Dan Toogood]

I realize this question isn't directly related to the usage of ccp4i, however in the past information provided to me on the ccp4bb has been excellent.

As such, I was wondering if anyone has had trouble with their SeMet proteins becoming insoluble upon IPTG induction.  The current protein I am working with has 'leaky' _expression_ and for the wild-type protein when it is not induced has more soluble protein than when it is.  However, in the case of the SeMet protein, upon induction with IPTG all SeMet protein becomes insoluble. I was wondering if anyone could provide some insights for experimental design as to how I can bypass the induction step or improve solubility with IPTG induction.

Thanks so much,

R. Daniel Toogood
MSc. Student
University of Saskatchewan
Saskatoon, Sask. Canada