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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 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.
A schematic representation of the pBR322 vector with restriction sites indicated in blue. pBR322 is a plasmid and was one of the first widely used E. coli cloning vectors . Created in 1977 in the laboratory of Herbert Boyer at the University of California, San Francisco , it was named after Francisco Bolivar Zapata , the postdoctoral researcher ...
Plasmid vectors minimalistically consist of an origin of replication that allows for semi-independent replication of the plasmid in the host. Plasmids are found widely in many bacteria, for example in Escherichia coli , but may also be found in a few eukaryotes, for example in yeast such as Saccharomyces cerevisiae . [ 8 ]
The entire set of fragments must be cloned together with the vector, and separation of clones can occur after. In either case, the fragments are ligated into a vector that has been digested with the same restriction enzyme. The vector containing the inserted fragments of genomic DNA can then be introduced into a host organism. [1]
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 ...
An example of a plasmid cloning vector which modifies the inserted protein is pFUSE-Fc plasmid. In order to genetically engineer insulin, the first step is to cut the MCS in the plasmid being used. [8] Once the MCS is cut, the gene for human insulin can be added making the plasmid genetically modified.
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]