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 11154070.
  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.
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