We are currently contemplating the acquisition of
an automated imaging system for crystallization screen plates (96-well).
Thanks to Zolt, Renaud and Catherine from Sanofi-Strasbourg, we bought
last year a Formulatrix Rock Imager.
The system is composed of a big box, containing your 96 wells
I have been trying to express a rat protein in bacteria. The MBP-fusion
expressed at very high level (~ 40 mg/L) while the GST-fusion and His-tag
only gave inclusion bodies. The problem is that all protein runs in the void
volume on a size-exclusion column (s-200, hepes, pH 7.4, 200 mM NaCl),
Hi,
You can use the on-line software EMBOSS.
You can use it at the following site :
http://bips.u-strasbg.fr/EMBOSS/
enjoy it !
Claudine
Jacob Keller wrote:
Dear Crystallographers,
Does anyone know of a bioinformatics counterpart of ccp4? It seems
like there should really be such an
I thought that as author of Scala I might put in my 2 penn'th to this
discussion, FWIW
1. I've never been able to find any useful distinction between Rsym
Rmerge, and when filling in the PDBs request for both (undefined by
them and irritatingly restricted to 0.99, at least in Autodep) I
A newer one for Ruby is (of course) BioRuby, which I use for simple
manipulation of sequences or chopping up structures.
It's quite nice for this, but I don't think it's as feature-rich as
Bio{Perl,Python,Java} though.
Hello,
We have some X-ray kit that is now surplus to requirements. We appreciate
that it is not state-of-the-art, but if anyone would like it they are very
welcome to come and pick it up:
The generator is a Nonius FR-591 system with a Enraf-Nonius DIP2000
detector. The MAC X-ray Optical
Dear all,
we are looking to crystallise some hydrophobic peptides in different
organic solvents, but find polystyrene plates are not sufficiently
resistent.
Also, it is not always obvious from which plastic plates are made. In
other words, anyone knows of:
- cleared polypropylene sitting
I wholly agree with the below. I am not sure how well E.coli can correctly fold
snaky
misfolded/unfolded protein that are chaperoned by folded tags! Not to rule out
that tags do it
sometimes...
Folded by association for insoluble proteins has often not worked well for
me. Sometimes, when it
Chen and David,
Before adding detergent, be forewarned that the MPB in many fusions
will not bind to an amylose column in the presence of most
detergents, particularly maltoside detergents. It has been the bane
to us so we have engineered MBP vectors with His tags to deal with
this.
Salt crystals are common in macromolecular crystallography. Has anyone
tried to tabulate salt crystal forms that commonly occur?
I just identified a salt crystal as Mirabilite, made of Na2SO4ยท10H2O.
The high water content makes them rather soft, and may not be recognized
as salt right away. In
Hi Chen,
Since you recognize that this is a protein expression problem, the best way
to get return of your investment of efforts is to get soluble expression
instead of trying to solubilize the protein down stream.
Many good suggestions have been proposed. I just want to add one more thing
Hi all
I am using COOT for model building and refinement.
i have to introducea ligand in my model
i have downloaded .cif file from RCSB.
However while importing the cif file i m getting the warning message of
having No restraints in the CIF file
is there any problem in the format of the following
Your cif-file contains coordinates, but as far as I could see it does not
contain bond lengths, and angles and their deviations. That is what coot
complains about.
Unless your cif-file describes a modified spermine you could use the
spermine already described in the refmac5 library:
In coot,
Hello all,
I am expecting a somewhat homogeneous reply to this one, but that is fine and
welcomed as are anecdotal experiences.
I am running co crystallisation experiments and have thus far been trying under
oil screens with some success (i.e. various hits in various conditions
resulting in
Hello Vineet,
This seems old one and doesn't have all restraints.
you may use refmac dictionary for spermine. I am sure this
will work in coot also. you may find it at
/ccp4-6.0.2/lib/data/monomers/s/SPM.cif
thanks
Manish
Vineet Gaur [EMAIL PROTECTED] wrote: Hi all
I am using COOT for
Don't call them co-crystals until you've actually shot them and seen
the density.
It will depend not only on the ligand but on the ligor(?) as well.
You don't mention the identity of your ligand or ligor(?). You mention
that some of your screens are under oil, so I will point out that some
I want to thank the very many people who responded to my inquiry. I'm
overwhelmed and really appreciative. I've still got to read through
it all, but a clear (near-unanimous) consensus is emerging.
All the best, and again thanks very much for the help. I really
appreciate it.
Bill
Brenda,
Generally speaking it is not abnormal to at least *hope* for co-crystals to
appear at or around the condition(s) where the native crystals are grown.
Therefore many people in fact begin looking for co-crystals by screening a
fine grid around the expected crystallization conditions for the
Dear All:
Recently I am pursuing the crystallziation of a complex formd by two
individual proteins and I met several interesting problems though they are
kind of off-topic.
Any suggestions for these problems will be highly appreciated.
BIAcore showed about
Hello Everyone,
When I refined a structure, I found strong difference density Fo-Fc at 3
sigma contour for the for five residues which already have occupancy of 1. The
density is continuous and so strong as if I did not put the residues there. Why
was that? Can I put greater than 1
Dear Joe,
working with divalent ions, I often get salt crystals (most surely
phosphate). I do not have any table available, but can give you the
commercial kits numbers going with it. Just let me know.
On Jan 22, 2008, at 6:12 PM, Joe Krahn wrote:
Salt crystals are common in
Dear Brenda,
I am running co crystallisation experiments and have thus far been
trying under
oil screens with some success (i.e. various hits in various conditions
resulting in crystal growth). These conditions have varied from
the native
condition up until now.
Do you already have the
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