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A multiple cloning site (MCS), also called a polylinker, is a short segment of DNA which contains many (up to ~20) restriction sites—a standard feature of engineered plasmids. [1] Restriction sites within an MCS are typically unique, occurring only once within a given plasmid.
Several databases exist for restriction sites and enzymes, of which the largest noncommercial database is REBASE. [5] [6] Recently, it has been shown that statistically significant nullomers (i.e. short absent motifs which are highly expected to exist) in virus genomes are restriction sites indicating that viruses have probably got rid of these motifs to facilitate invasion of bacterial hosts. [7]
For optimal use, plasmids that are commonly used for gene cloning are modified to include a short polylinker sequence (called the multiple cloning site, or MCS) rich in restriction recognition sequences. This allows flexibility when inserting gene fragments into the plasmid vector; restriction sites contained naturally within genes influence ...
A schematic representation of the pBR322 vector with restriction sites indicated in blue. pBR322 is a plasmid and was one of the first widely used E. coli cloning vectors . Created in 1977 in the laboratory of Herbert Boyer at the University of California, San Francisco , it was named after Francisco Bolivar Zapata , the postdoctoral researcher ...
The restriction sites in the MCS are first cleaved by restriction enzymes, then a PCR-amplified target gene also digested with the same enzymes is ligated into the vectors using DNA ligase. The target DNA sequence can be inserted into the vector in a specific direction if so desired. The restriction sites may be further used for sub-cloning ...
A restriction map is a map of known restriction sites within a sequence of DNA. Restriction mapping requires the use of restriction enzymes . In molecular biology , restriction maps are used as a reference to engineer plasmids or other relatively short pieces of DNA, and sometimes for longer genomic DNA.
A restriction enzyme or restriction endonuclease is a special type of biological macromolecule that functions as part of the "immune system" in bacteria.One special kind of restriction enzymes is the class of "homing endonucleases", these being present in all three domains of life, although their function seems to be very different from one domain to another.
The restriction modification system (RM system) is found in bacteria and archaea, and provides a defense against foreign DNA, such as that borne by bacteriophages.. Bacteria have restriction enzymes, also called restriction endonucleases, which cleave double-stranded DNA at specific points into fragments, which are then degraded further by other endonucleases.
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