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The partitioning system interacts with the septation apparatus to ensure that each daughter receives a copy of the plasmid. Many biotechnology applications utilize mutated plasmids that replicate to high copy number. For example, pBR322 is a medium copy number plasmid (~20 copies/cell) from which several high copy number cloning vectors (>100 ...
Generally, each molecule of plasmid diffuses randomly, so the probability of having a plasmid-less daughter cell is 2 1−N, where N is the number of copies. For instance, if there are 2 copies of a plasmid in a cell, there is 50% chance of having one plasmid-less daughter cell. However, high-copy number plasmids have a cost for the hosting cell.
Only the cells with the plasmid containing amp R will survive. The origin of replication (ori), is derived from the plasmid pMB1. [6] [1] pUC19 is a high copy number plasmid. [3] The high copy number is a result of the lack of the rop gene and a single point mutation in the ori of pMB1. [7] [8]
Many plasmids have high copy numbers, for example, pUC19 has a copy number of 500-700 copies per cell, [6] and high copy number is useful as it produces greater yield of recombinant plasmid for subsequent manipulation. However low-copy-number plasmids may be preferably used in certain circumstances, for example, when the protein from the cloned ...
Plasmid Vector. Recombinant DNA vectors function as carriers of the foreign DNA. Plasmids are small, closed-circular DNA molecules that exist from the chromosomes of their host. Their replication is to be under stringent control (low copy number) or relaxed (high copy number).
The plasmid with a high copy number origin of replication will generally have a higher transfection efficiency than one with a low copy number origin, using a plasmid with an origin of replication that is active in the host cell can lead to a higher transfection efficiency. [16]
The plasmid pSC101, a natural plasmid from Salmonella panama, [5] confers tetracycline resistance which allows for simpler screening process with antibiotic selection, but it is a low copy number plasmid which does not give a high yield of plasmid. Another plasmid, RSF 2124, which is a derivative of ColE1, confers ampicillin resistance but is ...
For example, a DNA sequence for a protein of interest could be cloned or subcloned into a high copy-number plasmid containing the lac (often LacUV5) promoter, which is then transformed into the bacterium E. coli. Addition of IPTG (a lactose analog) activates the lac promoter and causes the bacteria to express the protein of interest. [2]