Background
The origin of replication determines
the vector copy number, which could typically be in the range of
25–50 copies/cell if the expression vector is derived from the low-copy-number
plasmid pBR322, or between 150 and 200 copies/cell if derived from the
high-copy-number plasmid pUC. The copy number influences the plasmid stability,
i.e. the maintenance of the plasmid within the cells during cell division.
A positive effect of a high copy number is the greater stability of the
plasmid when the random partitioning occurs at cell division. On the other
hand, a high number of plasmids generally decreases the growth rate, thus
possibly allowing for cells with few plasmids to dominate the culture,
since they grow faster. There appears to be no significant advantage of
using higher-copy-number plasmids over pBR322 -based vectors in terms of
production yields.
The origin of replication also
determines the plasmid's compatibility: its ability to replicate
in conjunction with another plasmid within the same bacterial cell. Plasmids
that utilize the same replication system cannot co-exist in the same bacterial
cell. They are said to belong to the
same compatibility group. The introduction of a new
origin, in the form of a second plasmid from the same compatibility group,
mimics the result of replication of the resident plasmid. Thus any further
replication is prevented until after the two plasmids have been segregated
to different cells to create the correct prereplication copy number.
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(compatibility group) |
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pBR 322 and its derivatives (i.e, pET vectors) |
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pUC vectors |
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pACYC and its derivatives |
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pSC101 and its derivatives |
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ColE1 |
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Single-stranded DNA is synthesized. | ||
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For replication in eukaryotic cells |
Back to Origin of Replication related material Back to Vector Image
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