I am doing secretion of recombinant protein using E.coli. Before this, my protein was being secreted outside the cell into the medium and periplasm. When I want to repeat the same experiment after few months, the protein accumulated mostly in cytoplasm. How can I solve this problem? If this is because I am using old glycerol stock, I already retransformed to get freshly prepared transformants.
Shambhuprasad T.k.
Try inducing it with low amount of IPTG and grow the culture at 25 degree and induce with low IPTG overnight at 16-18 degree.
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