synthetic proteins

Cornelius Krasel krasel at alf.biochem.mpg.de
Tue Oct 18 11:18:41 EST 1994


lw75 (Lynne_A_WHITEHEAD at UMAIL.UMD.EDU) wrote:
: I have a question regarding the production of synthetic proteins.  I wanted
: to know in order to determine the various functions of different domains of
: a protein, if it's easier to produce synthetic proteins (with desired
: mutations) (obviously, also knowing the sequence) from "scratch"

By synthesizing the proteins chemically (only feasible for rather short
peptides, I don't know where a sensible limit is) or expressing the domains
in E. coli / Yeast / Sf9. You would always have to show that your domain
is functional.

If you don't need large amounts of protein, you could also try in vitro
transcription/translation.

: or doing controlled proteolytic digestion.  I assume that the second option is
: easier, but more difficult to control and more room for error.

You have to purify the original protein and have good control over the
digestion.

I would say it is dependent on the availability of the original protein
you want to study and the ease with which you can functionally express
(and purify) fragments of the protein in a foreign system. It also depends
on the number of fragments you want to make. For purification, inclusion
of a "tag" would probably be a good idea.

--Cornelius.

--
/* Cornelius Krasel, Abt. Lohse, Genzentrum, D-82152 Martinsried, Germany */
/* email: krasel at alf.biochem.mpg.de                 fax: +49 89 8578 3795 */
/* "People are DNA's way of making more DNA." (Edward O. Wilson, 1975)    */



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