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Dear Weijun Have a look at our REFOLD database - there are some examples of chaperone assisted refolding as well as many protocols for refolding of disulphide-rich proteins http://refold.med.monash.edu.au good luck ashley On 24/03/2007, at 12:23 PM, [log in to unmask] wrote: > Hi Weijun, > > you can get a kit from Takara (chaperone kit #3340) which has five > different chaperone plasmids (combinations of various chaperone). It > worked for me assofar that I could express huge amounts of the > chaperones, > but my target protein was either not expressed at all any more > (seems as > if E. coli was all busy expressing chaperons) or still insoluble ;-( > And unfortunately the structures of the chaperones were all solved > already > ;-) > > If disulfides might be the problem - try expression into the > periplasma > (some pET vectors encode leader sequences for that). > > Otherwise try other expression systems (baculovirus, mammalian....) > > All the best, Petra > > >> Dear All, >> >> I got enclusion body in many cases when I tried to express human >> proteins in E coli. I would like some suggestions on how to go about >> it. I would also like to try co-expression of GroEL/GroES or >> DnaJ/DnaK, and would like to know where to get the plasmids. >> >> Any help or comments would be appreciated. >> Weijun >> > > > -- > Petra Verdino, PhD > Research Associate > The Scripps Research Institute, BCC206 > 10550 North Torrey Pines Road > CA 92037 La Jolla, USA > tel:1-858-784-2294 > fax:1-858-784-2980 Ashley Buckle Ph.D NHMRC Senior Research Fellow The Department of Biochemistry and Molecular Biology School of Biomedical Sciences, Faculty of Medicine & Victorian Bioinformatics Consortium (VBC) Monash University, Clayton, Vic 3800 Australia http://www.med.monash.edu.au/biochem/staff/abuckle.html iChat/AIM: blindcaptaincat skype: ashley.buckle Tel: (613) 9905 3781 (office) Tel: (613) 9905 1653 (lab) Fax : (613) 9905 4699