Multiple cloning site
Encyclopedia
A multiple cloning site also called a polylinker, is a short segment of DNA
which contains many (up to ~20) restriction sites
- a standard feature of engineered plasmids. Restriction sites within an MCS are typically unique, occurring only once within a given plasmid. MCSs are commonly used during procedures involving molecular cloning or subcloning. Extremely useful in biotechnology
, bioengineering, and molecular genetics, MCSs let a biotechnologist insert a piece of DNA or several pieces of DNA into the region of the MCS. This can be used to create transgenic organisms, also known as genetically modified organisms (GMOs).
is pUC18. Its polylinker region is composed of several restriction enzyme recognition sites, that have been engineered into a single cluster (the polylinker). It has restriction sites for various restriction enzymes, including EcoRI
, BamHI
and PstI
. Another vector used in genetic engineering is pUC19
, which is similar to pUC18, but its polylinker region is reversed.
DNA
Deoxyribonucleic acid is a nucleic acid that contains the genetic instructions used in the development and functioning of all known living organisms . The DNA segments that carry this genetic information are called genes, but other DNA sequences have structural purposes, or are involved in...
which contains many (up to ~20) restriction sites
Restriction sites
Restriction sites, or restriction recognition sites, are locations on a DNA molecule containing specific sequences of nucleotides, which are recognized by restriction enzymes...
- a standard feature of engineered plasmids. Restriction sites within an MCS are typically unique, occurring only once within a given plasmid. MCSs are commonly used during procedures involving molecular cloning or subcloning. Extremely useful in biotechnology
Biotechnology
Biotechnology is a field of applied biology that involves the use of living organisms and bioprocesses in engineering, technology, medicine and other fields requiring bioproducts. Biotechnology also utilizes these products for manufacturing purpose...
, bioengineering, and molecular genetics, MCSs let a biotechnologist insert a piece of DNA or several pieces of DNA into the region of the MCS. This can be used to create transgenic organisms, also known as genetically modified organisms (GMOs).
The pUC18 and pUC19 polylinker
One bacterial plasmid used in genetic engineering as a plasmid cloning vectorCloning vector
A cloning vector is a small piece of DNA into which a foreign DNA fragment can be inserted. The insertion of the fragment into the cloning vector is carried out by treating the vehicle and the foreign DNA with a restriction enzyme that creates the same overhang, then ligating the fragments...
is pUC18. Its polylinker region is composed of several restriction enzyme recognition sites, that have been engineered into a single cluster (the polylinker). It has restriction sites for various restriction enzymes, including EcoRI
EcoRI
EcoRI is an endonuclease enzyme isolated from strains of E. coli, and is part of the restriction modification system.In molecular biology it is used as a restriction enzyme. It creates sticky ends with 5' end overhangs...
, BamHI
BamHI
BamHI is a restriction enzyme, derived from Bacillus amyloliquefaciens. It has the recognition site , and leaves a sticky end. One of the earlier enzymes to be used, it is popular for historical reasons, but also because digestion leaves a GATC overhang compatible with many other enzymes...
and PstI
PstI
PstI, is a Type II restriction endonuclease from Providencia stuartii. PstI recognition and cut site are as follows:...
. Another vector used in genetic engineering is pUC19
PUC19
pUC19 is one of a series of plasmid cloning vectors created by Messing and co-workers in the University of California. The p in its name stands for plasmid and UC represents the University in which it was created. It is a circular double stranded DNA and has 2686 base pairs...
, which is similar to pUC18, but its polylinker region is reversed.