NEB Impact protein purification system

Keith Jolley K.Jolley at soton.ac.uk
Mon Oct 27 05:18:25 EST 1997


So far I've got very low levels of expression of a membrane protein
(not expected to be soluble) cloned into both pCYB1 and pCYB3.  The
only strain where any expression was detectable by Western blotting
was BL21 - presumably due to the insoluble protein being broken down
in the others.  I was also getting some self-cleavage of the product
prior to attempting to purify it on a chitin column.  I tried the MBP
control and the levels of expression seen didn't match those suggested
in the protocol.
	Incidentally, I've since expressed the protein to much higher
levels using a T7 promoter and a 6-His tag (Invitrogen's Xpress
system).

On Fri, 24 Oct 1997 12:20:12 GMT, Michael.Vork at FYS.rulimburg.nl
(Michael  Vork) wrote:

>Hi there,
>
>
>Has anybody tried the new england biolabs impact system ? If so, please share 
>your experiences with me !
>
>Thanx 
>
>Michael Vork

-- 
Dr. Keith Jolley
Dept. of Molecular Microbiology
Southampton General Hospital, UK
Tel: 01703-798895  Fax: 01703-774316



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