How to avoid inclusion bodies ?

NZ nzheng at titan.oit.umass.edu
Tue May 23 11:07:14 EST 1995


I have a question related to this actually. The protein I am working on 
also goes to inclusion bodies. Recently, I become aware of the fact that 
some people are able to extract and solubilize their insoluble proteins 
from the inclusion bodies fraction by adding sarkosyl. Anyone has 
comments on this? Does it work in most cases?

Ning
Umass at Amherst

Clemens Vonrhein (vonrhein at bio5.chemie.uni-freiburg.de) wrote:
: 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
: ---

: ***************************************************************************
: * Clemens Vonrhein   email vonrhein at bio5.chemie.uni-freiburg.de           *
: *                    WWW   http://bio5.chemie.uni-freiburg.de/~vonrhein/  *
: *                                                                         *
: *         Institut f. Org. Chemie u. Biochemie                            *
: *         Albertstr.21                                                    *
: * D-79104 Freiburg i.Br. (Germany)                                        *
: *  Tel.: 761/203-6061   FAX: 761/203-5987                                 *
: ***************************************************************************



More information about the Proteins mailing list