Dear All,
I'd like to espress a protein for crystallisation as a whole, but also to try
separate expression of its two domains.
Although prediciting domain boundaires is not difficult, I know that it is
worthwhile testing several constructs to improve espression in soluble form. In
several works I've seen that the choice is to keep or give up some aminoacids
(for example 4 or 5) at the N and/or C-termini and to test them to see which
expresses best.
I was wondering if there are defined "rules" to choose these variants and/or
to modify the N and C termini of a protein of interest when it is not important
to keep them "true" (=native) in order to optimise expression levels.
Is there any good reference about this, please?
Personal experiences are welcome.
Many thanks,
Claudia
Claudia Scotti Dipartimento di Medicina Sperimentale Sezione di Patologia
Generale Universita' di Pavia Piazza Botta, 10 27100 Pavia Italia Tel. 0039
0382 986335/8/1 Facs 0039 0382 303673
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