GST-tagged protein purication from inclusion bodies

wattne via methods%40net.bio.net (by watnne from googlemail.com)
Tue Jan 12 12:18:31 EST 2010


On 12 Jan., 14:14, krishn pratap singh <kpsinghni... from gmail.com> wrote:
> Hi,
> Can anyone suggest me a method to purify my GST-tagged proteins from
> inclusion bodies. In which vector should i clone it to get the expression in
> soluble form.

Hi,
as GST is almost a protein on its own, it is already not only a means
for purification, but also for solubility. Usually it's difficult to
purify protein from inclusion bodies, particularly getting it out in
native or functional form; refolding is a pain and doesn't always
work. Therefore I would suggest preventing the protein to be put into
inclusion bodies, ie by reducing the expression level. Maybe either
lower temperature or less IPTG or what your inducer is, or shorter
expression time, or choosing a vector with weaker promotor.


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