Posted 14 March 2012 - 05:45 AM
I have cloned it in pGEX-6p1 and pET 28a. and expressed it but the expressed protein afer sonication when ran on SDS-PAGE is going in the pellet. so it seems that my protein is forming inclusion bodies. further I want to crystallize the protein. so I do not want to compromise with the concentration and structure of the protein. Please suggest me some cheap methods. ( the size of gene of interst is 966bp).
Posted 14 March 2012 - 10:07 AM
Posted 14 March 2012 - 10:27 AM
thanks for the reply. but i already did that.. i have decreased the induction time and IPTG concentration. cloned and expressed in both dh5 alpha and bl21 cells.. with 16 degree as my induction temp...
Induce less aggressively to lower the amount of protein being made. Culture your cells at lower temperature (25-30C, possibly lower).
Posted 25 April 2012 - 04:31 PM