A lot of your detection and purification systems are relying on the c terminal end. I reckon without doing an n-term fusion the only plausible possibilities are translation termination (really bad codons, cryptic terminators?) Or an obfuscated c terminal. Internal folding of the terminus might hide it from affinity chromatography or immunostaining?
I say try n-term fusion, then gene synthesis. Remember to include some useful system when synthesising, such as biobrick cassettes and a bunch of miscellaneous sites. If it's biobrick compatible you might even submit it to the registry.. Personally I recommend Mrgene.com by the way. They have very poor communication but they give an automatic quote while designing the gene and the design system is pretty intuitive. Oh, and they're pretty competitive. --- Twitter: @onetruecathal Sent from my beloved Android phone. On 26 Sep 2010 03:31, "Peter Ellis" <[email protected]> wrote: On 26/09/2010 02:13, DK wrote: > > In article<[email protected]>, Peter Ellis<[email protected]> wrote: >> >> >> A co... True, true, though at some point the distinction between "protein fragment" and "peptide" starts to become a little blurred! I'll bear this in mind when making antibodies in future. The other project I'm working on involves distinguishing between three protein families with >=90% amino acid identity. Good luck doing that without peptide ABs... but that's a whole different story. Not a new story if I'm honest, if you do a search on this newsgroup, it was a couple of years ago that I first started noticing I'd ended up with a particularly recalcitrant set of target genes. Peter _______________________________________________ Methods mailing list [email protected] http://www... _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
