Re: [ccp4bb] Using chaperones to boost expression in E. coli

2011-04-29 Thread Kothe, Michael
chael From: Olga Kolaj [mailto:olga.ko...@gmail.com] Sent: Tuesday, April 26, 2011 5:17 AM To: Kothe, Michael Cc: CCP4BB@jiscmail.ac.uk Subject: Re: [ccp4bb] Using chaperones to boost expression in E. coli Michael, We have put together a review about use of folding modulators to im

Re: [ccp4bb] Using chaperones to boost expression in E. coli

2011-04-26 Thread Olga Kolaj
Michael, We have put together a review about use of folding modulators to improve expression in E. coli some time ago which might be useful to you (Kolaj et al Microb Cell Fact. 2009 Jan 27;8:9). Here is the link: http://www.ncbi.nlm.nih.gov/pubmed/19173718

Re: [ccp4bb] Using chaperones to boost expression in E. coli

2011-04-22 Thread Shaun Lott
As with all these things, YMMV. In our experience, chaperone co-expression can help not at all, can produce more folded protein or can produce more soluble protein, with co-purifying chaperone. With GroEL/S at least, chaperone release is easy (add ATP) and sometimes results in the liberation of

Re: [ccp4bb] Using chaperones to boost expression in E. coli

2011-04-22 Thread aaleshin
Hi Michael, It worked for me with one viral enzyme but did not work with 2 others. In the successful case, I used Takara's chaperone plasmids to screen for the best composition of chaperones. The GroEL-GroES improved the yield of the soluble enzyme, but I got same activity (per 1L of media) as

Re: [ccp4bb] Using chaperones to boost expression in E. coli

2011-04-22 Thread Yoder, Marilyn
ical Sciences University of Missouri-Kansas City 5007 Rockhill Rd. Kansas City, MO 64110 Phone: 816-235-1986 From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Kothe, Michael Sent: Friday, April 22, 2011 10:28 AM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Using chaperones to b

[ccp4bb] Using chaperones to boost expression in E. coli

2011-04-22 Thread Kothe, Michael
Dear ccp4bb, I am curious to hear of examples where the expression of well-behaved protein was achieved by the coexpression of chaperones in E. coli. I know the appropriate strains and vectors exist, but I can't remember hearing of a successful case. I have heard anecdotally of several cases wh