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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]
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.
Addgene facilitates the exchange of genetic material between laboratories by offering plasmids and their associated cloning data to non-profit and academic laboratories around the world. Addgene provides a free online database of plasmid cloning information and references, including lists of commonly used vector backbones, popular lentiviral ...
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.
Baculovirus gene transfer into Mammalian cells (BacMam) is the use of a baculovirus to deliver genes to mammalian cells. [1] [2] Baculoviruses are insect viruses that are typically not capable of infecting mammalian cells; however, they can be modified to express proteins in mammalian cells.
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]
Molecular cloning takes advantage of the fact that the chemical structure of DNA is fundamentally the same in all living organisms. Therefore, if any segment of DNA from any organism is inserted into a DNA segment containing the molecular sequences required for DNA replication, and the resulting recombinant DNA is introduced into the organism from which the replication sequences were obtained ...