Restriction enzyme

Restriction enzyme glossary
Restriction
The cutting of DNA at specific sites
Enzyme
A protein that catalyzes a chemical reaction
Molecular recognition
Used by restriction enzymes to locate specific sequences of DNA on which to bind and subsequently cleave
Recognition sequence
The DNA sequence to which restriction enzymes bind
Restriction site
The site of the DNA sequence where it is cleaved by the restriction enzyme
Restriction fragment
A DNA fragment resulting from the cutting of a DNA strand by a restriction enzyme

A restriction enzyme, restriction endonuclease, REase, ENase or restrictase is an enzyme that cleaves DNA into fragments at or near specific recognition sites within molecules known as restriction sites.[1][2][3] Restriction enzymes are one class of the broader endonuclease group of enzymes. Restriction enzymes are commonly classified into five types, which differ in their structure and whether they cut their DNA substrate at their recognition site, or if the recognition and cleavage sites are separate from one another. To cut DNA, all restriction enzymes make two incisions, once through each sugar-phosphate backbone (i.e. each strand) of the DNA double helix.

These enzymes are found in bacteria and archaea and provide a defense mechanism against invading viruses.[4][5] Inside a prokaryote, the restriction enzymes selectively cut up foreign DNA in a process called restriction digestion; meanwhile, host DNA is protected by a modification enzyme (a methyltransferase) that modifies the prokaryotic DNA and blocks cleavage. Together, these two processes form the restriction modification system.[6]

More than 3,600 restriction endonucleases are known which represent over 250 different specificities.[7] Over 3,000 of these have been studied in detail, and more than 800 of these are available commercially.[8] These enzymes are routinely used for DNA modification in laboratories, and they are a vital tool in molecular cloning.[9][10][11]

  1. ^ Roberts RJ (November 1976). "Restriction endonucleases". CRC Critical Reviews in Biochemistry. 4 (2): 123–64. doi:10.3109/10409237609105456. PMID 795607.
  2. ^ Kessler C, Manta V (August 1990). "Specificity of restriction endonucleases and DNA modification methyltransferases a review (Edition 3)". Gene. 92 (1–2): 1–248. doi:10.1016/0378-1119(90)90486-B. PMID 2172084.
  3. ^ Pingoud A, Alves J, Geiger R (1993). "Chapter 8: Restriction Enzymes". In Burrell M (ed.). Enzymes of Molecular Biology. Methods of Molecular Biology. Vol. 16. Totowa, NJ: Humana Press. pp. 107–200. ISBN 0-89603-234-5.
  4. ^ Arber W, Linn S (1969). "DNA modification and restriction". Annual Review of Biochemistry. 38: 467–500. doi:10.1146/annurev.bi.38.070169.002343. PMID 4897066.
  5. ^ Krüger DH, Bickle TA (September 1983). "Bacteriophage survival: multiple mechanisms for avoiding the deoxyribonucleic acid restriction systems of their hosts". Microbiological Reviews. 47 (3): 345–60. doi:10.1128/MMBR.47.3.345-360.1983. PMC 281580. PMID 6314109.
  6. ^ Kobayashi I (September 2001). "Behavior of restriction-modification systems as selfish mobile elements and their impact on genome evolution". Nucleic Acids Research. 29 (18): 3742–56. doi:10.1093/nar/29.18.3742. PMC 55917. PMID 11557807.
  7. ^ Roberts RJ (April 2005). "How restriction enzymes became the workhorses of molecular biology". Proceedings of the National Academy of Sciences of the United States of America. 102 (17): 5905–8. Bibcode:2005PNAS..102.5905R. doi:10.1073/pnas.0500923102. PMC 1087929. PMID 15840723.
  8. ^ Roberts RJ, Vincze T, Posfai J, Macelis D (January 2007). "REBASE--enzymes and genes for DNA restriction and modification". Nucleic Acids Research. 35 (Database issue): D269-70. doi:10.1093/nar/gkl891. PMC 1899104. PMID 17202163.
  9. ^ Primrose SB, Old RW (1994). Principles of gene manipulation: an introduction to genetic engineering. Oxford: Blackwell Scientific. ISBN 0-632-03712-1.
  10. ^ Micklos DA, Bloom MV, Freyer GA (1996). Laboratory DNA science: an introduction to recombinant DNA techniques and methods of genome analysis. Menlo Park, Calif: Benjamin/Cummings Pub. Co. ISBN 0-8053-3040-2.
  11. ^ Massey A, Kreuzer H (2001). Recombinant DNA and Biotechnology: A Guide for Students. Washington, D.C.: ASM Press. ISBN 1-55581-176-0.