GST-fusion protein degrades in-vivo
Posted 18 May 2011 - 04:15 AM
Posted 19 May 2011 - 02:14 AM
Edited by Andrey, 19 May 2011 - 02:15 AM.
Posted 19 May 2011 - 11:57 AM
some say it is due to unspecific cleavage of the thrombin cleavage site in E. coli ...you can try using different strains like e.g. JM108/109.
you can use 1% glucose in the medium to prevent leaky expression of the tac promoter, additionally just do a short induction (not more than 2-3h) and use a protease inhibitor after lysing your cells!