What is polylinker sequence?
A polylinker is a short DNA sequence containing two or more different sites for cleavage by restriction enzymes. Polylinkers are introduced into vectors to make cloning easier by providing sites that allow cloning DNA, cut with any of a number of different restriction enzymes, into a single plasmid.
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: 1. The pMB1 replicon rep responsible for the replication of plasmid (source – plasmid pBR322).
What does pUC18 stand for?
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. It is a circular double stranded DNA and has 2686 base pairs.
What is the size of pUC18?
Thermo Scientific pUC18 vector is a small, high copy number, E. coli plasmid, 2686 bp in length. It contains identical multiple cloning site (MCS) as pUC19 vector except that it is arranged in opposite orientation.
Why is pUC18 used?
Vectors pUC18 and pUC19 are small high-copy number plasmids that are widely used for cloning and manipulation of DNA fragments (9).
What type of vector is pUC18?
pUC18 is a small, high copy cloning vector for replication in E. coli. It has been constructed using the ampicillin resistance gene and the pMB1 origin of replication from pBR322. The pMB1 of pUC18 differs from the pBR322 origin by a single point mutation and the lack of the rop gene, leading to a high copy number.
Is pUC18 and expression vector?
These vectors provide novel cloning possibilities in addition to simple, non-expensive, high level expression of recombinant proteins in E. coli. Vectors pUC18 and pUC19 are small high-copy number plasmids that are widely used for cloning and manipulation of DNA fragments (9).
What is the insertional inactivation?
Insertional inactivation is a technique used in recombinant DNA technology. In this procedure, a bacteria carrying recombinant plasmids or a fragment of foreign DNA is made to insert into a restriction site inside a gene to resist antibiotics, hence causing the gene to turn non-functional or in an inactivated state.
Why is insertional inactivation detected?
The presence of a chromogenic substrate gives blue coloured colonies in absence of an insert/in non-transformants presence of an insert in the enzyme site results into insertional inactivation of the α-galactosidase colonies which do not produce colour.
Is pUC18 ampicillin resistant?
The artificial plasmid pUC18 has been genetically engineered to include (1) a gene for antibiotic resistance to Ampicillin (amp R), and (2) a gene (and its promoter) for the enzyme beta-galactosidase (lacZ).