how to make a protein soluble
Hzhen at freeuk.com
Fri Mar 15 13:23:47 EST 2002
> I'm currently working with the pex7 gene in pet 28a vector.
> i ran an sds gel and found that the protein of interest is located in the
> insoluble portion of the cell.. my question is what are some of the ways I
> can make this protein soluble? Thank you.
It's a matter of luck whether you get soluble protein or not. As
mentioned by someone else, having a fusion partner sometimes helps with
solubility, the common ones are GST, thioredoxin, maltose-binding
protein (supposed to be best for solubility). Changing growth condition
also helps - you should aim to reduce to rate of protein expression,
expressing the protein too quickly can result in aggregation of
protein. Maybe chaperone can help, and if it has a physiological
partner which binds to it, perhaps coexpression with it may also be
useful. If this protein has binds to any ion or other prosthetic
groups, try adding it to the growth media.
You can also try refolding the protein if necessary, more work perhaps
but sometimes the only solution. If you are really unlucky, it may not
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