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Other cloning vectors include the pUC series of plasmids, and a large number of different cloning plasmid vectors are available. Many plasmids have high copy numbers, for example, pUC19 has a copy number of 500-700 copies per cell, [ 6 ] and high copy number is useful as it produces greater yield of recombinant plasmid for subsequent manipulation.
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]
A P1-derived artificial chromosome, or PAC, is a DNA construct derived from the DNA of P1 bacteriophages and Bacterial artificial chromosome.It can carry large amounts (about 100–300 kilobases) of other sequences for a variety of bioengineering purposes in bacteria.
For example, pBR322 is a medium copy number plasmid (~20 copies/cell) from which several high copy number cloning vectors (>100 copies/cell) have been derived by mutagenesis, such as the well known pUC series. [1] This delivers the convenience of high plasmid DNA yields but the additional burden of the high copy number restricts the plasmid size.
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 and Raymond L. Rodriguez. The p stands for "plasmid," and BR for "Bolivar" and "Rodriguez."
There are many types of cloning vectors such as plasmids and phages. In order to carry out recombination between vector and the foreign DNA, it is necessary the vector and DNA to be cloned by digestion , ligase the foreign DNA into the vector with the enzyme DNA ligase .
Insertional mutagenesis is when transposons function as vectors to help remove and integrate genetic sequences. Given their relatively simple design and inherent ability to move DNA sequences, transposons are highly compatible at transducing genetic material, making them ideal genetic tools.
The loading capacity of cosmids varies depending on the size of the vector itself but usually lies around 40–45 kb. The cloning procedure involves the generation of two vector arms which are then joined to the foreign DNA. Selection against wild type cosmid DNA is simply done via size exclusion.
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