Re: [ccp4bb] off topic

Hi Anamika, Proteins with high sequence identity can behave very differently when expressed in bacteria. Sumo (or other tags) may or may not help. If it does, you can express your SH2 with a tag and remove the tag after purification if the tag is a problem for functional studies. The

Re: [ccp4bb] off topic

Hi Anamika, I guess you tried several growth conditions. You might try to do a heat shock growth. Grow at 37C until OD 0.6, then move the bacteria to 42C for half hour, then to 30C and grow them for 20 hours. Another strategy is to add just before induction 1,2 or 3% Ethanol. Not sure these will

Re: [ccp4bb] off topic

Hi Anamika, I did a search and it looks like researchers are using either mammalian cells or baculovirus to express this protein. I don't have experience with this particular construct so could you tell me why you choose *E. coli*? I run a protein expression facility and we typically use HEK or

Re: [ccp4bb] off topic

Hey Anamika I know SH2 domain is very much well-studied domain, and i am not sure why are you facing troubles in the expression of this protein. Just go through the literature and read about different protocol of expression of SH2 domain from several different proteins.. Well, reading that you

Re: [ccp4bb] ccp4i2 crashes on start

One possible source of the problem that we've come across before... ensure the follow environment variable isn't set: QT_PLUGIN_PATH Best wishes, Tom On 4 July 2017 at 14:55, Stuart McNicholas wrote: > Dear Johannes, >Many apologies, this behaviour should

[ccp4bb] ARP/wARP webservice

Dear Colleagues, We are happy to announce a redesigned and extended webservice for remote ARP/wARP execution. In addition to the crystallographic protein chain tracing, many other functionalities of ARP/wARP are now also available online. These include: - classic protein model building

Re: [ccp4bb] REBATCH error mystery

REBATCH is old and I can’t remember how it works :-( > On 6 Jul 2017, at 15:18, Graeme Winter wrote: > > HI Phil, > > Yes, probably. However it’s still a mystery I would like to understand. Also, > I use REBATCH for lots of things not just trimming data sets (i.e.

Re: [ccp4bb] REBATCH error mystery

HI Phil, Yes, probably. However it’s still a mystery I would like to understand. Also, I use REBATCH for lots of things not just trimming data sets (i.e. manually updating BATCH numbers) and - this is the real fun one - if I rerun the processing with 900 images REBATCH works fine! … following

[ccp4bb] off topic

Hi, Is anyone has worked with STAT1 proteins? I have cloned the SH2 domain of STAT1 protein into pet28a vector but there was no expression so far or rather say inconsistent expression. Sometimes the expression was in inclusion bodies. I have tried different methods to pull out the protein from

Re: [ccp4bb] REBATCH error mystery

REBATCH should generally be replaceable by Pointless I think Latest latest Pointless (prerelease update 043, version 1.11.3) removes batch headers for leading or trailing batch numbers which are not present in the reflection list (for you, Graeme) > On 6 Jul 2017, at 15:04, Graeme Winter

[ccp4bb] Scientific Software Developer position at RCSB Protein Data Bank (RCSB.org) at the University of California San Diego (UCSD)

For contact information and other positions, please see http://www.rcsb.org/pdb/static.do?p=general_information/about_pdb/contact/job_listings.html Scientific Software Developer Develop, implement,

[ccp4bb] REBATCH error mystery

Afternoon all, Technical harking back to the 90’s FORTRAN CCP4 question if I may I have an MTZ file which has no spots on first couple of images, so the batch headers at the start exist but there are no BATCH values which correspond to these in the actual reflection data I have been bashing

[ccp4bb] RELEASE NOTE SIMPLE 2.5

Dear All: We announce the release of a new version of our program package SIMPLE (v2.5) for single-particle cryo-EM *ab initio* 3D reconstruction (web: simplecryoem.com) New features include: * A new DDD movie pre-processing program that implements motion correction based the same principal