Hi everyone
I have 2 vectors and I want to transform these into one bacteria cell (SHuffle strain).
Do you think co-transformation (Transforming 2 vectors together in one step into the competent cell) has an effect on Protein expression?
Or, that I have to transform the vectors step by step (e.g. transforming vector number 1 after that competent this cell and transform vector number 2).
is really different between these two strategies for Protein expression?
Note: I used co-transformation for these 2 vectors But the Expression rate was so low Compared with when I Expressed one vector Separately.
If there are toxicity issues with one plasmid that is ameliorated by the other plasmid, then it is possible that cotransformation is not a good idea. Since the second plasmid won't have time to express the protecting protein to make the first plasmid not toxic. Therefore you might be selecting for non-toxic (non-expressing) variants.
On the other hand, if there are no toxicity issues then I can't really think of a reason for your results.
Hi Dr. Michael J. Benedik
Thank you for your comment.
My proteins are not toxic to each other.
Do you think if one of my proteins is toxic, secondary protein expression But transfer to the inclusion body?
I doubt it has anything to do with propagation of inclusion bodies. But if transforming sequentially works, then it may not be worth worrying about. Just stick with what you know works. It is faster to do the 2 steps transformation than to find out your culture is not exprssing protein.
Dr. Michael J. Benedik
Thank you so much for your comment.
actually, My results show no difference between CO-transformation and competent in protein expression (I competented My Bacteria and Used step-by-step transformation).
However, thank you.
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