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A plasmid is a small, extrachromosomal DNA molecule within a cell that is physically separated from chromosomal DNA and can replicate independently. They are most commonly found as small circular, double-stranded DNA molecules in bacteria; however, plasmids are sometimes present in archaea and eukaryotic organisms.
F-prime cell contains F-plasmid that integrates with the chromosomal DNA and carries part of the chromosomal DNA along with it while being excised from the chromosome. Thus F-prime plasmid is the plasmid, containing part of the chromosomal DNA which can be transferred to recipient cell, along with the plasmid during conjugation. [4]
conjugative - mediate DNA transfer through conjugation and therefore spread rapidly among the bacterial cells of a population; e.g., F plasmid, many R and some col plasmids. nonconjugative - do not mediate DNA through conjugation, e.g., many R and col plasmids. The pBR322 plasmid is one of the first plasmids widely used as a cloning vector.
A plasmid partition system is a mechanism that ensures the stable inheritance of plasmids during bacterial cell division. Each plasmid has its independent replication system which controls the number of copies of the plasmid in a cell. The higher the copy number, the more likely the two daughter cells will contain the plasmid.
The plasmid therefore requires a selectable marker such that those cells without the plasmid may be killed or have their growth arrested. Antibiotic resistance is the most commonly used marker for prokaryotes. The transforming plasmid contains a gene that confers resistance to an antibiotic that the bacteria are otherwise sensitive to.
Once the MCS is cut, the gene for human insulin can be added making the plasmid genetically modified. After that, the genetically modified plasmid is put into the bacterial host and allowed to divide. To make the large supply that is demanded, the host cells are put into a large fermentation tank that is an optimal environment for the host.
The total number of a particular plasmid within a cell is referred to as the copy number and can range from as few as two copies per cell to as many as several hundred copies per cell. [11] Circular bacterial plasmids are classified according to the special functions that the genes encoded on the plasmid provide.
An origin of transfer – A plasmid with no origin of transfer is non-mobilizable. [2] The transfer genes – Though a functioning set of tra genes is necessary for plasmid transfer, they may be located in a variety of places including the plasmid in question, another plasmid in the same host cell, or even in the bacterial genome. [3]