pRSET/histag expression problems

Drobert2 at utk.edu Drobert2 at utk.edu
Wed Jun 12 15:14:59 EST 1996


I have a problem with the expression of a membrane protein in E. coli using 
the histag/pRSET system.  We are using the protease deficient strain BL21lysE. 
 The expression level is low (0.2 mg/liter culture) and much of the protein 
does not contain a histag (apparently it is cleaved off during expression or 
subsequent purification).  The protein does not accumulate in inlcusion bodies 
and apparently is inserted ok into membranes.  Has anyone else seen these 
problems and could offer advice on how to improve expression/stability.



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