Trying to express recombinant GST-proteins after overnight culture at 18ºC with 0.1 mM IPTG I observed also with my fusion protein a band corresponding to beta-lactamase after maldi analysis. How can I eliminate this expression?
HI there,
You are using an AmpR plasmid so either you do as Stefan suggested or you may purify your fusion protein on GSH agarose: beta-lactamase will be excluded from the column and your fusion recovered with GSH elution (in theory...).
Hi,
Thanks all, I was thingking that maybe my ampicillin could be old?, or to express for shorter time. I expressed already other constructs but it is the first time I had so much betalactamase.
You might have put too much ampicillin in your culture which provokes the increase of the copy number of AmpR plasmid per cell (and so beta-lactamase activity is also increased)
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