What is the MCS shown on the plasmid map?
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. Restriction sites within an MCS are typically unique, occurring only once within a given plasmid.
What is a pUC19 plasmid?
pUC19 is a commonly used plasmid cloning vector in E. The molecule is a double-stranded circular DNA (2686 base pairs in length). The pUC19 plasmid confers ampicillin resistance and complement defects in β-galactosidase in appropriate host strains.
Where is pUC19 from?
pUC19 is one of a series of plasmid cloning vectors created by Joachim Messing and co-workers. The designation “pUC” is derived from the classical “p” prefix (denoting “plasmid”) and the abbreviation for the University of California, where early work on the plasmid series had been conducted.
What is the host of plasmid pUC19?
Mutants will be selected for by screening for the insertional inactivation of the α-complementation shared by pUC19 and its host DH5α. The pUC19 plasmid (Fig. 1) will be linearized in the MCS, and a fragment 1.7kb in length will be removed from the pBR322 plasmid (Fig. 2) by restriction digests.
What is the advantage of having an MCS in a plasmid?
A multiple cloning site (MCS, or Polylinker region) is a DNA region within a Plasmid that contains multiple unique Restriction enzyme cut sites. Plasmids are very useful in biotechnology and one key feature of their use is the multiple cloning site, which allows for foreign DNA to be inserted into the plasmid.
Is pUC19 a plasmid?
Thermo Scientific pUC19 vector is a small, high copy number, E. coli plasmid, 2686 bp in length. It contains identical multiple cloning site (MCS) as pUC18 vector except that it is arranged in opposite orientation.
What is the difference between pUC18 and pUC19?
pUC19 is identical to pUC18 except that they contain multiple cloning sites (MCS) arranged in opposite orientations. pUC18/19 plasmids contain: The pMB1 replicon rep responsible for the replication of plasmid (source – plasmid pBR322).
Does plasmids have selectable marker?
The antibiotic resistance gene allows for selection in bacteria. However, many plasmids also have selectable markers for use in other cell types. A short single-stranded DNA sequence used as an initiation point for PCR amplification or sequencing. Primers can be exploited for sequence verification of plasmids.
What is the advantage of using polylinker in a vector?
Plasmid vectors that contain a polylinker will be cut only once by multiple restriction enzymes, each acting at its own site. Inclusion of a polylinker in a plasmid vector thus permits cloning of restriction fragments generated by cleavage of DNA with multiple different restriction enzymes.
Where is the multiple cloning site ( MCS ) located in pUC19?
The multiple cloning site (MCS) region is split into codons 6-7 of the lacZ gene, providing for many restriction endonucleases restriction sites. In addition to β-galactosidase, pUC19 also encodes for an ampicillin resistance gene (amp R ), via a β-lactamase enzyme that functions by degrading ampicillin and reducing its toxicity to the host.
Where does the name PUC come from in plasmid cloning?
pUC19 is one of a series of plasmid cloning vectors created by Joachim Messing and co-workers. The designation “pUC” is derived from the classical “p” prefix (denoting “plasmid”) and the abbreviation for the University of California, where early work on the plasmid series had been conducted.
How is the MCS reversed in pUC18?
Standard E. coli vector with a multiple cloning site (MCS) for DNA cloning. The MCS is reversed in pUC18. To see this sequence with restriction sites, features, and translations, please download SnapGene or the free SnapGene Viewer.
Where does the resistance gene pUC19 come from?
In addition to β-galactosidase, pUC19 also encodes for an ampicillin resistance gene (amp R), via a β-lactamase enzyme that functions by degrading ampicillin and reducing its toxicity to the host. The ori site, or origin of replication, is derived from the plasmid pMB1.