Unexpressable protein

Cathal Garvey via methods%40net.bio.net (by cathalgarvey from gmail.com)
Sun Sep 26 08:57:55 EST 2010


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" <pjie2 from cam.ac.uk> wrote:

On 26/09/2010 02:13, DK wrote:

>
> In article<8g7dvnFjcgU1 from mid.individual.net>, Peter Ellis<pjie2 from cam.ac.uk>
 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


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