I am having problem with expression of gene. I got 2 clone, my first clone is 369 bp which is well expressed. When i made another clone by adding 63 bp of positive charged amino acid with my first clone, i cant expressed it. I am using DE3 cell for expression and my vector is pRSETA which has his tag protein.
I tried in different condition like temperature and time of induction by IPTG. Could you give me any suggestion for this.