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| Hi there, Have you ever got any answer to your questions about the pUC and pBR322 you posted in 1999? If so, could you please share them with me? Thanks a lot. Mark I have a question reagarding the copy number of pBR322 based plasmids. I have looked for references in the literature, but cannot find the specific answer I seek. I know that the main differences between pBR322 and pUC18 (with respect to replication and copy number) is that pUC18 was created from pBR322 by deletion of the rop and mutatiion in the ori. From Current Protocols and Maniatis, I have found that the rop is responsible for keeping the plasmid replication from going "nuts". As a consequence, the copy number of pBR322 is ~10 copies per cell while the copy number of pUC18 is ~700 copies per cell. The mutation in the ori is also a contributing factor in this higher copy number. I would like to know what the expected copy number would be of pBR322 WITHOUT the rop but with no mutation in the ori? I would expect that it would be much greater than pBR322 but less than pUC18. How much of a role does the mutation in ori play? Idea, answers, and references would be greatly appreciated!! Thanks! Chuck Miller |
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