On 12 Jan., 14:14, krishn pratap singh <[email protected]> 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. _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
