Re: [ccp4bb] What can a learning student should post in our forum- are any restrictions for discussing science[what is science?] with intellectuals :)

2009-02-27 Thread Kevin Cowtan
Jayashankar wrote: I am still confused with what is right and wrong in this world of research where the point of reference changes with time and condition. Some body made me to accept that science and philosophy are verymuch different?Are they ? No,

[ccp4bb] Off topic: Coot and shared libraries

2009-02-27 Thread Partha Chakrabarti
Hi, I am running a 64 bit Fedora 10 machine and having problems with coot, if i say ldd coot-real linux-gate.so.1 = (0x5000) libguilegtk-2.0.so.0 = not found libgthread-2.0.so.0 = /lib/libgthread-2.0.so.0 (0x00df8000) librt.so.1 = /lib/librt.so.1 (0x005f) libguile.so.17

Re: [ccp4bb] Off topic: Coot and shared libraries

2009-02-27 Thread Tim Gruene
Coot sets up the paths before it starts. That's why you don't call coot-real but the script coot which modifies LD_LIBRARY_PATH and some other environment variables before actually calling the binary. Therefore applying ldd to coot-real is not telling you much unless you set LD_LIBRARY_PATH

Re: [ccp4bb] Off topic: Coot and shared libraries

2009-02-27 Thread Paul Emsley
Partha Chakrabarti wrote: Hi, I am running a 64 bit Fedora 10 machine and having problems with coot, if i say ldd coot-real linux-gate.so.1 = (0x5000) libguilegtk-2.0.so.0 = not found libgthread-2.0.so.0 = /lib/libgthread-2.0.so.0 (0x00df8000) librt.so.1 = /lib/librt.so.1

Re: [ccp4bb] Off topic: Mammalian gene expression in E. coli

2009-02-27 Thread Stephen Weeks
Just in case anyone else encounters the same problem, like Artem I have had a few SUMO fusion constructs that have stubbornly refused to cleave even with a 1mg of enzyme (Ratio ~ 1:100) at 37c in the presence of low concentrations of chaotrope. In all cases the problem was solved by inserting

[ccp4bb] Refmac5 - high res (1.2A) refinement problem

2009-02-27 Thread Michael Weyand
Dear experts, here is a Refmac question (or at least a refinement question): the case: data set was collected at the synchrotron with high intensity or better high total dose. Data set scales to 1.2A resolution. Refinement with refmac5_0066, using hydrogens, anisotropic B-factors, automatic

Re: [ccp4bb] Refmac5 - high res (1.2A) refinement problem

2009-02-27 Thread George M. Sheldrick
It would be possible to model all of that with SHELXL, but it is a complex problem and would need time and patience with any program. One advantage of SHELXL would be the flexible refinement of occupancies for the different conformations. George Prof. George M. Sheldrick FRS Dept. Structural

Re: [ccp4bb] Off topic: Coot and shared libraries

2009-02-27 Thread Frances C. Bernstein
I can't help on software but this would explain Gordon Brown's presence: http://www.google.com/hostednews/ukpress/article/ALeqM5jQKS8GcXtpz_FvAK08ybc5QRfRBQ Isn't google amazing? Frances =

Re: [ccp4bb] Off topic: Mammalian gene expression in E. coli

2009-02-27 Thread artem
Hi, In a case like the one Raji outlined below - after all the attempts - I would have most cerainly switched to insect cells as the next step :) If you suspect that protein of interest has large disordered regions, expression in a higher order system by itself may not be enough (still good

[ccp4bb] In-house X-ray systems: what is your experience

2009-02-27 Thread Sergei Strelkov
Dear All, following a good discussion recently on the crystallization imaging systems, I wanted to start a discussion with respect to choosing a new in-house X-ray source (rotating anode). Bruker is there with Microstar-H and Microstar Ultra. Rigaku has 007HF and FR-E+. I wondered if people

Re: [ccp4bb] Off topic: Mammalian gene expression in E. coli

2009-02-27 Thread Narayanan Ramasubbu
ar...@xtals.org wrote: Hi, In a case like the one Raji outlined below - after all the attempts - I would have most cerainly switched to insect cells as the next step :) If you suspect that protein of interest has large disordered regions, expression in a higher order system by itself may not

Re: [ccp4bb] In-house X-ray systems: what is your experience

2009-02-27 Thread VSS Fairweather, Science 02
Dear Sergei, We have recently had a Microstar installed from Bruker. It had some serious teething problems and it took close to a year from the point of installation in our building until it could be used for any data collection. Many parts of the original machine delivered had to be replaced

Re: [ccp4bb] Refmac5 - high res (1.2A) refinement problem

2009-02-27 Thread Paul Swepston
Michael: I attended the CCP4 study weekend in January and there were many discussions about radiation damage. In your case I wonder if recollecting the data with a lower over all dosage might get rid of the problem. Either collect with an attenuated beam or collect the data twice as fast.

[ccp4bb] radiation damage

2009-02-27 Thread James Holton
As much as I hate to see the CCP4BB used for shameless self-promotion of articles, I did just publish a Beginner's Guide to Radiation Damage: http://journals.iucr.org/s/issues/2009/02/00/issconts.html My answers to Paul and Michael's posts would be cut-and-pasted from it, so I think it

Re: [ccp4bb] radiation damage

2009-02-27 Thread Gerard DVD Kleywegt
I even made the cover! And I hope that (after looking at the cover) all you BBers will forgive me just this once as this was clearly a once-in-a-lifetime opportunity. Or, at least, I HOPE only once. Congratulations, James. But surely you could have found a better example of medical imaging?

Re: [ccp4bb] Off topic: Mammalian gene expression in E. coli

2009-02-27 Thread Artem Evdokimov
Hello, The short answer is 'yes'. If you can use both methods :) The issue with limited proteolysis lies in the questionable state of the full-length protein - if the stuff is nasty and misfolded, then fagments generated by proteolytic digest aren't going to be meaningful. On the other hand if

Re: [ccp4bb] Off topic: Mammalian gene expression in E. coli

2009-02-27 Thread Ho-Leung Ng
Most of the poorly cleavable fusion proteins (usually MBP-TEV) that I've seen turned out to be solubly aggregated. ho UC Berkeley -- Date:Fri, 27 Feb 2009 07:23:43 -0500 From:Stephen Weeks stephen.we...@verizon.net Subject: Re: Off topic: Mammalian gene

Re: [ccp4bb] radiation damage

2009-02-27 Thread James Holton
Gerard DVD Kleywegt wrote: Congratulations, James. But surely you could have found a better example of medical imaging? For instance, here: http://www.ibeatyou.com/competition/867945/craziest-x-ray/entry/be24b3/coke-bottle --dvd Unfortunately, (unlike the infamous coke bottle image),