Smallest mutatable catalitic or fluorescent protein
AEVDOKIMOZ at cinci.rr.com
Thu Nov 28 00:36:23 EST 2002
"EK" <khatipovNO-SPAM at NO-SPAMuchicago.edu> wrote in message
news:AReF9.18$N4.4037 at news.uchicago.edu...
> I would like to construct the following fusion:
> Peptide1 ---Carrier(6His)protein ---Peptide2
A few suggestions:
1) auto-biotynilation can be accomplished by using a specific peptide
sequence that gets biotinylated (you would need to augment expression of the
relevant transferase, otherwise biotinylation won't be complete). So pretty
much anything, any small useful protein, can be made into biotinylated one
2) as far as activity etc. is concerned - there is a multitude of choices.
If you like phosphatase assays (e.g. PNPP) - express one of the small acidic
tyrosine phosphatases, e.g. the bovine one, which is tiny, nicely folded, is
easily produced in e. coli, and the structure's known. If you fancy other
assays - just let us know which assay you like best, and I have no doubt
that the group will come up with suitable candidates :) There's a relatively
new red fluorescent protein from a sea anemone, not a GFP relative, which I
think is reasonably small, too.
3) Beware of proteolysis. Many of the library sequences will get chewed up
in E. coli, so you'll never get a complete library.
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