[ccp4bb] Jeff Christensen is out of the office.

I will be out of the office starting 06/05/2009 and will not return until 06/08/2009. I will respond to your message when I return. Thanks!

Re: [ccp4bb] TEV nucleotude sequence with restriction site

Does anybody have a TEV-protease-site-coding nucleotide sequence with a commonly-used restriction site in it, preferably right at the end? Alternatively, does some somebody know of a program to determine all equivalent codon permutations for a small coding region, filtered for resulting restric

Re: [ccp4bb] TEV nucleotude sequence with restriction site

Try type II restriction enzymes. They cleave outside of their recognition sites. So you can use one vector for different inserts. There may be a type I enzyme site for TEV site, I haven't found a good one yet. It's likely some inserts will have the same site internally, making the vector less usef

Re: [ccp4bb] TEV nucleotude sequence with restriction site

Hi Jacob, If you're including the TEV site in your primer, then you won't need a restriction site after it (unless you're planning to recycle the vector afterwards for other inserts). Having the protease site too close to the protein can often make it difficult to cut. Cheers, Charlie Quot

Re: [ccp4bb] images

Hi Sean Ash Buckle has already developed one! Tools for general deposition will be released shortly. http://tardis.edu.au/ Cheers J Sean Seaver wrote: > I started a poll to find out whether crystallographers need and are > interested in an X-ray diffraction data bank. Will crystallographers

Re: [ccp4bb] TEV nucleotude sequence with restriction site

I checked out the Sheffield et al paper, and the restriction sites there are all just after the TEV site, thereby including, as Cynthia mentioned, at least an extra H beyond the obligatory G from the TEV site. I was hoping to be able to have only the G. (Since I am cloning in the TEV site with m

Re: [ccp4bb] TEV nucleotude sequence with restriction site

I'm not quite sure what you want, but I have a series of vectors encoding various N-terminal tags and fusions, all followed by a TEV site. They have an MCS standard to many pET vectors. Therefore, they are designed to clone your gene in using the NdeI site at the 5' end (which will, after

Re: [ccp4bb] TEV nucleotude sequence with restriction site

You seem to be describing the MCS found in many TEV-site-containing expression plasmids (am I missing something?) E.g., look at the sequences in Sheffield et al., Protein Expression and Purification 15, 34 –39 (1999) (let me know if you want a PDF, I don't want to send it to the whole bb)

[ccp4bb] TEV nucleotude sequence with restriction site

Dear Crystallographers, Does anybody have a TEV-protease-site-coding nucleotide sequence with a commonly-used restriction site in it, preferably right at the end? Alternatively, does some somebody know of a program to determine all equivalent codon permutations for a small coding region, filte

Re: [ccp4bb] images

I started a poll to find out whether crystallographers need and are interested in an X-ray diffraction data bank. Will crystallographers find this resource helpful and be willing to submit their structures? I hope you will take a moment to share your opinion via the poll and/or by posting any que

Re: [ccp4bb] Refmac specific questions

Hi Partha, It should be depended on the input model : MIXEd Some atoms with isotropic, some with anisotropic B-values. In this case input file (PDB) defines which atom should be refined isotropicly and which anisotropicly. The atoms with ANISOU card are refined anisotropicly. ta l

[ccp4bb] Refmac specific questions

Hi, Sorry to sound stupid, could someone explain what does refmac do if one chooses "mixed (bref MIXED)" or "overall (bref OVER)" B factor refinement in the context of TLS refinement? Is it something like: Isotropic for the main chain and/or waters and anisotropic for the side chain? Regards, Pa