Restriction Enzymes


Type II restriction enzymes are the molecular scissors that catalyse the double‐strand cleavage of deoxyribonucleic acid (DNA) at specific base sequences. They are essential tools for manipulating DNA including but not limited to cloning, analysis and sequencing. Recent advances have made it possible to design and engineer chimaeric nucleases to target specific DNA sites within a genome thus making them useful tools to carry out gene therapy.

Keywords: restriction enzymes; endonucleases; methylases; restriction–modification; recombinant DNA; chimaeric nucleases

Figure 1.

Representation of multimodular enzymes. (a) FokI (a type IIs restriction enzyme); (b) I‐TevI (a homing endonuclease). R, recognition domain; EN, endonuclease domain.

Figure 2.

Representation of various chimaeric nucleases. ZF‐EN, Ubx‐EN and Gal4‐EN were made by fusing the isolated nuclease domain (EN) of FokI to the zinc‐finger motif, helix–turn–helix motif and Gal4, respectively. R, recognition domain; EN, endonuclease domain.



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Further Reading

Aggarwal AK and Wah DA (1998) Novel site‐specific DNA endonuclease. Current Opinion in Structural Biology 8(1): 19–25.

Berg JM (1995) Zinc finger domains: from prediction to design. Accounts of Chemical Research 28: 14–19.

Bibikova M, Beumer K, Trautman JK and Carroll D (2003) Enhancing gene targeting with designed zinc finger nucleases. Science 300: 764.

Bibikova M, Carroll D, Segal DJ et al. (2001) Stimulation of homologous recombination through targeted cleavage by chimeric nucleases. Molecular and Cellular Biology 21: 289–297.

Bibikova M, Golic M, Golic KG and Carroll D (2002) Targeted chromosomal cleavage and mutagenesis in Drosophila using zinc finger nucleases. Genetics 161: 1169–1175.

Carroll D (1996) Homologous genetic recombination in Xenopus: mechanisms and implications for gene manipulation. Progress in Nucleic Acid Research and Molecular Biology 54: 101–125.

Choo Y and Klug A (1997) Physical basis of a protein–DNA recognition code. Current Opinion in Structural Biology 7: 117–125.

Durai S, Mani M, Kandavelou K et al. (2005) Zinc finger nucleases: custom‐designed molecular scissors for genome engineering of plant and mammalian cells. Nucleic Acids Research 33: 5978–5990.

Gormley NA, Watson MA and Halford SE (2005) Bacterial restriction–modification systems,

Kandavelou K, Mani M, Durai S and Chandrasegaran S (2004) Engineering and applications of chimeric nucleases. In: Pingoud AM (ed.) Nucleic Acids and Molecular Biology, vol. 14, pp. 413–434. Berlin: Springer‐Verlag.

Kandavelou K, Mani M, Durai S and Chandrasegaran S (2005) “Magic” scissors for genome surgery. Nature Biotechnology 23: 686–687.

Kandavelou K, Mani M, Reddy SPM and Chandrasegaran S (2005) Ligation: theory and practice,

Kim J‐S and Pabo CO (1998) Getting a handhold on DNA: design of poly zinc finger proteins with femtomolar dissociation constants. Proceedings of the National Academy of Sciences of the USA 95: 2812–2817.

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New England Biolabs (2000–2001) New England Biolabs Catalog and Technical Reference. Beverly, MA: New England Biolabs.

Porteus MH and Baltimore D (2003) Chimeric nucleases stimulate gene targeting in human cells. Science 300: 763.

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Williams RJ (2001) Restriction endonucleases and their uses. Methods in Molecular Biology 160: 409–429.

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How to Cite close
Mani, Mala, Kandavelou, Karthikeyan, and Chandrasegaran, Srinivasan(Dec 2007) Restriction Enzymes. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1002/9780470015902.a0000973.pub3]