Nicking enzyme

A nicking enzyme (or nicking endonuclease) is an enzyme that cuts one strand of a double-stranded DNA at a specific recognition nucleotide sequences known as a restriction site. Such enzymes hydrolyse (cut) only one strand of the DNA duplex, to produce DNA molecules that are “nicked”, rather than cleaved.[1][2]

They can be used for strand-displacement amplification,[3] Nicking Enzyme Amplification Reaction, exonucleotyic degradation, the creation of small gaps,[4] or nick translation.[5] The latter process has been successfully used to incorporate both radioactively labelled nucleotides and fluorescent nucleotides allowing specific regions on a double stranded DNA to be studied.[5][6] Over 200 nicking enzymes have been studied, and 13 of these are available commercially[7] and are routinely used for research and in commercial products.

References

  1. Ando T, et. Al. (July 1969). "Isolation and characterization of enzymes with nicking action from phage T4-infected Escherichia coli". J. Biochem. 66 (1): 1–10. PMID 4309718.
  2. Morgan RD, Kong H, et al. (November 2000). "Characterization of the specific DNA nicking activity of restriction endonuclease N.BstNBI". Biol. Chem. 381 (11): 1123–5. doi:10.1515/BC.2000.137. PMID 1154070.
  3. Walker GT, Little MC, Nadeau JG, Shank DD. (Jan 1992). "Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system". Proc. Natl. Acad. Sci. U.S.A. 89 (1): 392–6. doi:10.1073/pnas.89.1.392. PMC 48243. PMID 1309614.
  4. Wang H, Hays JB. (October 2001). "Simple and rapid preparation of gapped plasmid DNA for incorporation of oligomers containing specific DNA lesions". Mol. Biotechnol. 19 (2): 133–40. doi:10.1385/MB:19:2:133. PMID 11725483.
  5. 1 2 Rigby PW, Dieckmann M, Rhodes C, Berg P (June 1977). "Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I". J. Mol. Biol. 113 (1): 237–51. doi:10.1016/0022-2836(77)90052-3. PMID 881736.
  6. Jo K, Dhingra DM, Odijk T,de Pablo JJ, Graham MD, Runnheim R, Forrest D, Schwartz DC (2007). "A single-molecule barcoding system using nanoslits for DNA analysis". Proc. Natl. Acad. Sci. U.S.A. 104 (8): 2673–8. doi:10.1073/pnas.0611151104. PMC 1815240. PMID 17296933.
  7. "REBASE Enzymes". Encyclopedia of restriction and nicking enzymes. Retrieved 2009-06-01.

External links

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