How to avoid inclusion bodies ?

[Clemens Vonrhein]

The subject says it all: I want to avoid the expression of a protein
as inclusion bodies.

As far as I read the literature, there are some rules of thumb:

- lower temperature (30, 25, 20, 15 degrees Celsius ?)
- low IPTG concentration
- rich medium ( terific broth, ...)
- high OD and short incubation times (OD=1.0-2.0 and t=1-2h)
- adding needed cofactors (FAD, FMN, ...)

Are these rules true in general ? Is there a good and recent review ?
What are your own experiences ?

Is it worth trying to get soluble protein ? Or is it better to get a 
lot of protein as inclusion bodies and try to refold it ?

BTW, I use a pET-22/BL21 system.

Any hints/answers/tips .. highly apreciated.

Thanks

Clemens
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* Clemens Vonrhein   email vonrhein at bio5.chemie.uni-freiburg.de           *
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