His tag increasing expression and solubility?!
Posted 06 May 2010 - 07:22 AM
So I usually get helpful advice on this forum, so here goes!
1) I expressed my gene with a his tag in BL21(DE3) at 30 degrees with 1mM IPTG. = Saw a good expression band on the SDS-page at 3 hours post induction with majority being soluble.
2) I expressed the same gene but without his tag in BL21(DE3) at 30 degrees with 1mM IPTG = Saw no exression whatsoever in both soluble and insoluble fractions.
What is going on? I am aware of his tag causing insolubility problems, but is it possible that a his tag is beneficial for this protein from an expression/solubility sense?
Attachment 1 : Gene 1 with tag. showing expression in the middle lanes(mid gel).
Attachment 2 : Gene 1 without tag showing no expression
Posted 13 May 2010 - 04:54 AM
Posted 13 May 2010 - 06:40 AM
Maybe you can indicate what band on the gel is your "overexpressed" gene?
Posted 13 May 2010 - 04:05 PM