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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 T7 expression system is used in the field of microbiology to clone recombinant DNA using strains of E. coli. [1] It is the most popular system for expressing recombinant proteins in E. coli. [2] By 2021, this system had been described in over 220,000 research publications. [3]
Usually the ultimate aim of expression cloning is to produce large quantities of specific proteins.To this end, a bacterial expression clone may include a ribosome binding site (Shine-Dalgarno sequence) to enhance translation of the gene of interest's mRNA, a transcription termination sequence, or, in eukaryotes, specific sequences to promote the post-translational modification of the protein ...
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]
The cloning vector may be DNA taken from a virus, the cell of a higher organism, or it may be the plasmid of a bacterium. The vector contains features that allow for the convenient insertion of a DNA fragment into the vector or its removal from the vector, for example through the presence of restriction sites.
The vector itself generally carries a DNA sequence that consists of an insert (in this case the transgene) and a larger sequence that serves as the "backbone" of the vector. The purpose of a vector which transfers genetic information to another cell is typically to isolate, multiply, or express the insert in the target cell.
An expression cassette is a distinct component of vector DNA consisting of a gene and regulatory sequence to be expressed by a transfected cell. [1] In each successful transformation, the expression cassette directs the cell's machinery to make RNA and protein(s). Some expression cassettes are designed for modular cloning of protein-encoding ...
A pUC19 cloning vector showing the multiple cloning site sequence with restriction enzyme sites. 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. [1]