RFI:Increasing amount of soluble protein in bacterial expression

Kevin Brady rcxkb at unix.ccc.nottingham.ac.uk
Mon Apr 2 12:32:45 EST 2001


Without killing the cells, the only way of getting proteins into the
media is getting them into the periplasmic space first (usually by
encoding a signal peptide at the start of your gene), and then the
protein may leach out of the cell by mechanisms that are not really
understood. This is not usually very efficient at all and you may be
better off just going for cytoplasmic or inclusion body purification. If
you're desperate to use media fractions, you might want to think about
using yeast e.g. Pichia pastoris. Invitrogen do the kit (no affil.)
which comes with vectors for intra and extra-cellular expression. P.
pastoris is well documented (though I haven't got references on me) for
secreting proteins in large amounts.
Best of luck
Kevin

--
Dr Kevin Brady
Organic Chemistry Section
School of Chemistry
University of Nottingham
Nottingham
UK
NG7 2RD
#################################################
Email : Kevin.Brady at nottingham.ac.uk or
        kevin at basil.chem.nott.ac.uk
URL   : http://www.nottingham.ac.uk/~pcxsc/thomas
#################################################


-------------- next part --------------
An HTML attachment was scrubbed...
URL: http://iubio.bio.indiana.edu/bionet/mm/methods/attachments/20010402/75111a41/attachment.html


More information about the Methods mailing list