Good vector for protein expression in bacteria
legatek at mcmail.cis.mcmaster.ca
Wed Jul 10 14:36:16 EST 2002
On 9 Jul 2002, Jonny Yokosawa wrote:
> I am planning on expressing a protein in bacteria, but I don't want it as a
> fusion protein, like the GST fusion proteins. I am considering adding 6xHis
> to the sequence, though. The multiple cloning site of the vectors I looked
> is located after the GST sequence (pGEX). I wonder if any of you would have
> any suggestions for a vector that I could use and if the 6xHis would be
> enough for purification purposes.
Invitrogen have a set of vectors designated pRSET that have a 6xHis tag
and an EK cleavage site, if that's important to you. They are under T7
control and work fine in BL21(DE3) in my experience. 6xHis is good for one
step of your purification, but you probably want to also include ion
exchange or gel filtration as well.
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legatek at mcmaster.ca Kyle Legate legatek at hotmail.com
Tower of Tongues:Thursday PM:10:30-11:30 EDT:http://cfmu.mcmaster.ca
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