Re: [ccp4bb] Dell-Apple Mac and graphics glasses

2008-11-03 Thread Christopher Bahl
You are right, that particular video card does not support stereo graphics. However, with a different card, they should still be able to make it work with that computer. http://reald-corporate.com/scientific/graphics_cards_by_os.asp We were able to pick up an ATI FireGL V7100 for about $130

[ccp4bb] Semet in non-auxotrophic strains

2008-11-03 Thread Jacob Keller
Dear Crystallographers, I have been using an E coli strain called Tuner (DE3) pLacI, which is not auxotrophic for methionine, nor is there an auxotrophic analogue available. It may be that my protein can be expressed in other strains, but this one works well right now, and it is usually

Re: [ccp4bb] Semet in non-auxotrophic strains

2008-11-03 Thread Guenter Fritz
Hi Jacob, you can express SeMet labeled protein in almost any E.coli expression strain if you use a defined minimal medium. Here is a receipe that worked quite well in our hands: http://strucbio.biologie.uni-konstanz.de/ccp4wiki/index.php/Expression_of_SeMet_labeled_proteins HTH Guenter Dear

Re: [ccp4bb] Semet in non-auxotrophic strains

2008-11-03 Thread Cynthia Kinsland
You can express Se-Met protein in non-auxotrophs using metabolic inhibition (Doublie. Methods Enzymol. 276, 523-530, 1997 and Van Duyne et al. J. Mol. Biol. 229, 105-124, 1993). The Studier auto-induction paper (Studier, Prot. Exp. Purif. 41, 207-234, 2005) describes an auto-induction

Re: [ccp4bb] Cryoprotectant for protein-DNA complex crystal

2008-11-03 Thread Phoebe Rice
I think we still have better luck with longer, slower, more gentle soaks - but its crystal-dependent. Try raising the [PEG] at the same time as you raise the [glycerol]. Phoebe Original message Date: Fri, 31 Oct 2008 18:57:53 -0400 From: Artem Evdokimov [EMAIL PROTECTED] Subject:

Re: [ccp4bb] Semet in non-auxotrophic strains

2008-11-03 Thread Jacob Keller
Dear Crystallographers, I am overwhlemed with all of the responses--thank you all very much for your help. This listserve is a fantastic resource, and it is great that there is patience for all of the mundane methods questions! I will try to do my part to help out when I can. Thanks, Jacob

[ccp4bb] unsubscribe ccp4bb

2008-11-03 Thread Vatsan Raman

Re: [ccp4bb] Semet in non-auxotrophic strains

2008-11-03 Thread Artem Evdokimov
Jacob, We routinely express SeMet proteins in native E. coli without any trouble. The method is a derivative of Doublie's. If you want, I would be more than happy to send you a protocol (we've modified the process to gain biomass yield). Artem -Original Message- From: CCP4 bulletin

[ccp4bb] m-3m vs. m-3

2008-11-03 Thread yanming Zhang
Dear 'old' crystallographers, During one case of structure solution, the data processing programs output incorrect space group-primitive cubic P4132, which later found out that the correct one should be face centerd cubic f23. This problem was caused by perfect twinning. Now I'd like to

Re: [ccp4bb] Semet in non-auxotrophic strains (summary)

2008-11-03 Thread Artem Evdokimov
http://www.xtals.org/pdfs/selenium.pdf (sorry, for some reason first email went out w/o the hyperlink) Artem -Original Message- From: Artem Evdokimov [mailto:[EMAIL PROTECTED] Sent: Monday, November 03, 2008 9:26 PM To: 'Jacob Keller'; 'CCP4BB@JISCMAIL.AC.UK' Subject: RE: [ccp4bb]

Re: [ccp4bb] Semet in non-auxotrophic strains (summary)

2008-11-03 Thread Vangelis Christodoulou
While staying on the subject, I found the following paper very useful when trying to produce Se-Met labeled protein in HEK293 Flp-In. Barton et al. Highly efficient selenomethionine labeling of recombinant proteins produced in mammalian cells. Protein Sci (2006) vol. 15 (8) pp. 2008-13