I have a low copy plasmid (ori=pBR322) and I wanted change it to high copy (like ori=pUC).
This paper seemed to say that only one mutation was required to increase the copy number:
Article High copy number of the pUC plasmid result from a Rom/Rop su...
However, I made this mutation in my plasmid and it didn't change the copy number. My plasmid does not appear to have rop either.
What are the key mutations between pBR322 and pUC that cause pUC to have such a high copy number?
Are you sure it doesn't have rop, and it's not just present but unannotated? Origins of replication and the areas around them are often annotated really poorly because it's inconsequential information for most researchers.
If you are certain that's not the case then I think you would have to give the actual sequence of the plasmid for anyone to really speculate what's going on.
Hi Alexandra Johnson
I think I uploaded the plasmid map in this post. The mutation I made is annotated as "SNP for high copy pMB1." Previously, I was detecting rop by trying to align to a rop sequence I got from another plasmid map. I saw no alignment (forward or backwards), so I concluded rop was absent. Thank you very much.
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