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A restriction map is a map of known restriction sites within a sequence of DNA. Restriction mapping requires the use of restriction enzymes . In molecular biology , restriction maps are used as a reference to engineer plasmids or other relatively short pieces of DNA, and sometimes for longer genomic DNA.
Optical mapping [1] is a technique for constructing ordered, genome-wide, high-resolution restriction maps from single, stained molecules of DNA, called "optical maps". By mapping the location of restriction enzyme sites along the unknown DNA of an organism, the spectrum of resulting DNA fragments collectively serves as a unique "fingerprint" or "barcode" for that sequence.
One bacterial plasmid used in genetic engineering as a plasmid cloning vector is pUC18. Its polylinker region is composed of several restriction enzyme recognition sites, that have been engineered into a single cluster (the polylinker). It has restriction sites for various restriction enzymes, including EcoRI, BamHI, and PstI.
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 ...
conjugative - mediate DNA transfer through conjugation and therefore spread rapidly among the bacterial cells of a population; e.g., F plasmid, many R and some col plasmids. nonconjugative - do not mediate DNA through conjugation, e.g., many R and col plasmids. The pBR322 plasmid is one of the first plasmids widely used as a cloning vector.
Long-range restriction mapping is an alternative genomic mapping technique to short-range, also called fine-scale mapping. Both forms utilize restriction enzymes in order to decipher the previously unknown order of DNA segments; the main difference between the two being the amount of DNA that comprises the final map. The unknown DNA is broken ...
Cells which have been successfully transformed with pUC19 can be differentiated from cells which have not by growing them on media with ampicillin. Only the cells with the plasmid containing amp R will survive. The origin of replication (ori), is derived from the plasmid pMB1. [6] [1] pUC19 is a high copy number plasmid. [3]
Several databases exist for restriction sites and enzymes, of which the largest noncommercial database is REBASE. [5] [6] Recently, it has been shown that statistically significant nullomers (i.e. short absent motifs which are highly expected to exist) in virus genomes are restriction sites indicating that viruses have probably got rid of these motifs to facilitate invasion of bacterial hosts. [7]