Plasmids are circular double stranded DNA (dsDNA) molecules separated from cell chromosome DNA. These extra chromosomal DNA naturally exist in bacteria, yeast and some advanced eukaryotic cells, which have parasitic or symbiotic relationship with their host cells. The size of the plasmid ranges from several thousand base pairs to more than 100 kb. DNA fragments of up to 20 kb base pairs can be inserted into the plasmid vector. When the recombinant plasmid is transformed into E.coli cells, all antibiotic resistant progenies from the original transformed cells will contain plasmids with the same inserted DNA sequence. The inserted DNA replicates with the rest of the plasmid DNA and is separated into daughter cells as the colony grows. Some bacterial plasmids encode enzymes that inactivate antibiotics. Such resistant plasmids have become a major problem in the treatment of many common bacterial pathogens. To deal with the spread of resistant plasmids is an important challenge of modern medicine. So plasmid cloning is very important in such research of plasmids, and PCR is a common method for plasmid cloning.
Fig. 1 Schematic representation of plasmid cloning by PCR. (Qi R, et al. 2019)
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