A nicking enzyme (or nicking endonuclease) is an enzyme that cuts one strand of a double-stranded DNA or RNA[1] at a specific recognition nucleotide sequences known as a restriction site. Such enzymes hydrolyze (cut) only one strand of the DNA duplex, to produce DNA molecules that are “nicked”, rather than cleaved.[2][3]
They can be used for strand-displacement amplification,[4] Nicking Enzyme Amplification Reaction, exonucleolytic degradation, the creation of small gaps,[5] or nick translation.[6] 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.[6][7] Over 200 nicking enzymes have been studied, and 13 of these are available commercially[8] and are routinely used for research and in commercial products.
References
edit- ^ Wang X, Zhu B (22 May 2024). "SARS-CoV-2 nsp15 preferentially degrades AU-rich dsRNA via its dsRNA nickase activity". Nucleic Acids Research. 52 (9): 5257–5272. doi:10.1093/nar/gkae290. PMC 11109939. PMID 38634805.
- ^ 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. doi:10.1093/oxfordjournals.jbchem.a129107. PMID 4309718.
- ^ 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. S2CID 22472698.
- ^ 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. Bibcode:1992PNAS...89..392W. doi:10.1073/pnas.89.1.392. PMC 48243. PMID 1309614.
- ^ 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. S2CID 22156627.
- ^ a b 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.
- ^ 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. Bibcode:2007PNAS..104.2673J. doi:10.1073/pnas.0611151104. PMC 1815240. PMID 17296933.
- ^ "REBASE Enzymes". Encyclopedia of restriction and nicking enzymes. Retrieved 2009-06-01.