What is pUC19 plasmid used for?
pUC19 is a commonly used cloning vector that conveys the Amp resistance. The molecule is a small double-stranded circle, 2686 base pairs in length, and has a high copy number.
What is pUC19 plasmid?
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 are the features of pUC19 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 purpose of ampicillin resistance gene in this cloning vector?
In order to be useful in DNA cloning, however, a plasmid vector must contain a selectable gene, most commonly a drug-resistance gene encoding an enzyme that inactivates a specific antibiotic. As we’ve seen, the ampicillin-resistance gene (ampr) encodes β-lactamase, which inactivates the antibiotic ampicillin.
Why is pUC19 a great cloning vector?
Why is PUC19 a great cloning vector? pUC19 carries a 54 base-pair multiple cloning site polylinker that contains unique sites for 13 different hexanucleotide-specific restriction endonuclease.; vector and foreign DNA cut with the enzyme.
What is the difference between pBR322 and pUC19?
pUC19 has a smaller plasmid size in comparison to pBR322, which are 2686bp and 4361bp respectively (1). When present individually in a cell, pUC19 has a plasmid copy number of around 75 per cell at 37⁰C and >200 per cell at 42⁰C, whereas pBR322 has a plasmid copy number of only around 20 per cell (7, 4).
What are features of pUC19 vector?
The molecule is a small double-stranded circle, 2686 base pairs in length, and has a high copy number. pUC19 carries a 54 base-pair multiple cloning site polylinker that contains unique sites for 13 different hexanucleotide-specific restriction endonucleases (1).
What is the purpose of ampicillin resistance gene?
The ampicillin-resistance gene allows us to select which of the E. coli cells have been transformed based on their ability to grow in an environment that contains the antibiotic ampicillin.
What is ampicillin resistance?
Ampicillin resistance was defined as a minimum inhibitory concentration (MIC) greater than 16 g/mL.
How is pUC19 constructed?
pUC19 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 pUC19 differs from the pBR322 origin by a single point mutation and the lack of the rop gene, leading to a high copy number.
What is insertional activation?
Definition. Insertional activation/inactivation refers to either activation of an endogenous gene which is located near an integrated transgene, or to disruption of a gene or other functional sequence by insertion of a transposable element.