Star activity is the relaxation or alteration of the specificity of restriction enzyme mediated cleavage of DNA that can occur under reaction conditions that differ significantly from those optimal for the enzyme. The result is typically cleavage at non-canonical recognition sites, or sometimes complete loss of specificity.
Differences which can lead to star include low ionic strength, high pH, and high (> 5% v/v) glycerol concentrations.[1] The latter condition is of particular practical interest, since commercial restriction enzymes are usually supplied in a buffer containing a substantial amount of glycerol (50% v/v is typical), meaning insufficient dilution of the enzyme solution can cause star activity; this problem most often arises during double or multiple digests. Star activity can happen because of presence of Mg2+, as is seen in HindIII, for example.
The term star activity was introduced by Mayer[2] who characterized the modified activity in EcoRI.
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Star activity
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Transcription
External links
- Star Activity - New England Biolabs
- Star Activity (Relaxation of Specificity) - Fermentas
- Star activity of restriction enzymes - a detailed list from TaKaRa
- Wei, H; Therrien, C; Blanchard, A; Guan, S; Zhu, Z (2008). "The Fidelity Index provides a systematic quantitation of star activity of DNA restriction endonucleases". Nucleic Acids Research. 36 (9): e50. doi:10.1093/nar/gkn182. PMC 2396408. PMID 18413342.
References
- ^ Robinson, CR; Sliger, SG (1993). "Molecular Recognition Mediated by Bound Water : A Mechanism for Star Activity of the Restriction Endonuclease EcoRI". Journal of Molecular Biology. 234 (2): 302–306. doi:10.1006/jmbi.1993.1586. PMID 8230215.
- ^ Mayer, Hubert (1978). "Optimization of the EcoRI*-activity of EcoRI endonuclease". FEBS Lett. 90 (2): 341–44. doi:10.1016/0014-5793(78)80400-1. PMID 352725.
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This page was last edited on 24 May 2024, at 21:44