Dear ALL;
We are going to use the prescission protease cutting site (LEVLFQ/GP
) in our cloning vector to remove the His6 tag.
Do we need to insert some linkers between this cutting site and the
target protein to improve the cleavage efficiency?
Or we are
Dear McCully,
I always used this protease with a cleavage success near to 100% at 4°C over
night without shaking.
Concerning the construct, I putted the first Met just after the sequence
LEVLFQ/GP SSP, so I added only three aminoacids after the cleavage site.
All the best,
Marco
Il
Dear Jerry,
Our in-house series of vectors encode the Rhinovirus 3C-protease site, followed
directly by a NdeI site, which after cleavage leaves just GPHM on the front of
your protein. We routinely get 100% cleavage with incubation overnight at 4˚C
— and have several xtal structures to boot.
