TEV protease

Dr. Artem Evdokimov eudokima at mail.ncifcrf.gov
Sat Aug 11 18:56:13 EST 2001


It depends on what application do you need your TEV for. We use TEV for
most of our work, depending on the purificaiton scheme, MBP-Tev-Arg5 is
probably the most useful, though just plain old TEV-Arg5 or TEV-His6 is
also OK.

I do not know about TEV from Doudna lab, but all our TEV's are perfectly
soluble. What we do is a) express it as an MBP fusion (with
self-processing linker!) and b) use lower temperature for expression
(26-30C, not 37!). If you express any TEV at 37, it does, indeed,
misfold.

My boss (David Waugh) routinely sends out all sorts of TEV plasmids, all
you need to do is to fill the government paperwork out.

Some references:

1) Expression, purification, refolding, and characterization of
recombinant human interleukin-13: utilization of intracellular
processing.
Eisenmesser EZ, Kapust RB, Nawrocki JP, Mazzulla MJ, Pannell LK, Waugh
DS, Byrd RA.
Protein Expr Purif 2000 Nov;20(2):186-95

2) Kapust RB, Waugh DS.
Controlled intracellular processing of fusion proteins by TEV protease.
Protein Expr Purif. 2000 Jul;19(2):312-8.

In general, search Pubmed for 'WAUGH DS' and the references will pop up.
Or write him and I am sure he'll be able to send you the PDF files.

Artem

> And which one is, in relative terms, "the best"? (Or most frequently used).
> 6His-TEV mutant from Doudna is in inclusion bodies. What about yours?
> Is there a ref. to any of them or all?
> 
> Thanks,
> 
>         Dima

-- 
|Dr. Artem Evdokimov   Protein Engineering |
| NCI-Frederick        Tel. (301)846-5401  |
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|        eudokima at mail.ncifcrf.gov         |
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