L Bruce Sk lbrucesk at
Tue Mar 14 21:38:16 EST 1995

>In article <iert237-030395125450 at>,
iert237 at (zac ) says:
>        We are working with the GST-Fusion system and are having an
>difficult time getting a fusion product that is not degraded.  We have
>looked at almost every parameter, including induction time, culture
>sonication/lysis conditions, you name it.  We were also told some
>constructs just do no work well do to unknown problems linked with the
>of the protein we have cloned in.  We have now made a new construct that
>totally different in size (much larger but encoding the same initial
>region) and have found that degredation is so bad that we barely have any
>fusion product.  If anyone can help, I would GREATLY appreciate it. 
>we are using the pGex-2T vector
>Darren Boehning
>Thomas Jefferson University
>boehnin1 at
>I have had such problems too, and I found an easy way to have less of
>products by growing the cells in LB medium WITHOUT GLUCOSE, for only
>hours after induction with IPTG, if necessary you can also try to grow
the cells at
>30c after the induction (that with very usefull in my hands); finally I
would strongly
>recommand you to switch of E. coli strain to try a Omp minus.
>Good luck

Pharmacia can supply you with a 'GST Gene Fusion System' Troubleshooting
and Procedural Guide, now in its second edition for no charge.  Contact
your Pharmacia rep or the company directly to obtain (800) 526-3593

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