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An expression vector, otherwise known as an expression construct, is usually a plasmid or virus designed for gene expression in cells. The vector is used to introduce a specific gene into a target cell, and can commandeer the cell's mechanism for protein synthesis to produce the protein encoded by the gene.
The purpose of an MCS in a plasmid is to allow a piece of DNA to be inserted into that region. [2] An MCS is found in a variety of vectors, including cloning vectors to increase the number of copies of target DNA, and in expression vectors to create a protein product. [3] In expression vectors, the MCS is located downstream of the promoter. [2]
Vector map of pUC19. pUC19 is one of a series of plasmid cloning vectors designed by Joachim Messing and co-workers. [1] 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. [2]
E. coli is one of the most widely used expression hosts, and DNA is normally introduced in a plasmid expression vector. The techniques for overexpression in E. coli are well developed and work by increasing the number of copies of the gene or increasing the binding strength of the promoter region so assisting transcription. [3]
The CAG promoter is a strong synthetic promoter frequently used to drive high levels of gene expression in mammalian expression vectors. [1] [2] CAG promoter was constructed in the lab of Dr Jun-ichi Miyazaki [3] [4] from the following sequences: (C) the cytomegalovirus (CMV) early enhancer element,
A version called mPB was created by optimizing codon usage for mammalian (mouse) with a 20x increase in activity, [9] and further mutation screening generated hyPB with 10x the activity of mPB. [10] PiggyBac system have been successfully employed to express large genetic sequences, such as a doxycicline-inducible CRISPR interference system.
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