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Star Activity of Restriction Enzymes

Some restriction enzymes may cleave different base sequence than their recognition sequence, under sub-optimal reaction conditions. These conditions generally include high salt concentrations, impure templates, or a large excess of enzyme compared to substrate DNA. This reduced specificity is called Star Activity. The table below lists enzymes for which star activity may be observed together with the experimental conditions required to induce star activity. The alternative base sequences cleaved are noted where data are available.

Restriction enzyme Nomal recognition
sequence
Reaction
conditions*
Recognition
sequence**
Reference
Aat IIGACGT↓CE 22
Aor 13H IT↓CCGGAA, C, E 22
Ava IC↓YCGRGA, E 1
Ava IIG↓GWCCA, E 22
Bam H IG↓GATCCA, B, E, FGRATCC
GGNTCC
GGANCC
GGATYC
1,2,3,4
Ban IIGRGCY↓CA, E, F 22
Bgl IGCCNNNN↓NGGCF 22
Bgl IIA↓GATCTE 22
Bme T110 IC↓YCGRGE 
Bsp T104 ITT↓CGAAA, E 22
Bst P IG↓GTNACCA, F 22
Bst 1107 IGTA↓TACF 22
Dde IC↓TNAGA, B, E  
Eam 1105 IGACNNN↓NNGTCA, C, F 22
Eco O65 IG↓GTNACCE, F 22
Eco R IG↓AATTCA, B, E, FNAATTN4,8,9,10,11
Eco R VGAT↓ATCE RATATC
GNTATC
GANATC
GATNTC
GATANC
GATATY
12,22
Eco T22 IATGCA↓TF, H 22
Fba IT↓GATCAA, C, E, F 22
Hae IIIGG↓CCA 1
Hha IGCG↓CA, E 4
Hin c IIGTY↓RACE 22
Hin d IIIA↓AGCTTB, E RAGCTT
ANGCTT
AAKCTT
AAGMTT
AAGCNT
AAGCTY
10,13,22
Hpa IGTT↓AACA, E 1,22
Kpn IGGTAC↓CE 22
Mun IC↓AATTGA, F 22
Nco IC↓CATGGA, E 22
Nhe IG↓CTAGCA, C, E, F 22
Psp 1406 IAA↓CGTTE 22
Pst ICTGCA↓GA, E 1,4,22
Pvu IICAG↓CTGA, E NAGCTG
CNGCTG
CANCTG
CAGNTG
CAGCNG
CAGCTN
15,16,22
Sac IGAGCT↓CA, E 22
Sal IG↓TCGACA, E 4,22
Sau 3A I↓GATCA, E SATC
GMTC
GAKC
GATS
17,22
Sca IAGT↓ACTB, C, F 21
Sfi IGGCCNNNN↓NGGCCB, E 22
Smi IATTT↓AAATA, B, C, E 22
Spe IA↓CTAGTD, F 22
Sse 8387 ICCTGCA↓GGD, F 22
Ssp IAAT↓ATTA, C, E, F 22
Taq IT↓CGAA, C, F 22
Tth 111 IGACN↓NNGTCB, C, G NACNNNGTC
GNCNNNGTC
GANNNNGTC
GACNNNNTC
GACNNNGNC
GACNNNGTN
19
Van 91 ICCANNNN↓NTGGA 22
Vpa K11B IG↓GWCCA, C, E, F 22
Xba IT↓CTAGAA, E 1,4

*Reaction Conditions

A: high levels of glycerol
B: in the presence of Mn2+
C: alkaline pH
D: in the presence of DMSO
E: low ionic strength
F: high ionic strength;
G: in the presence of 2-mercaptoethanol

**Key

M=A or C; K=G or T; N=A, C, G, or T;
R=A or G; Y=C or T; W=A or T; S=G or C

References

  1. Nath, K. and Azzolina, B. A. (1981) Gene Amplif. Anal. 1:113-30.
  2. George, J., et al. (1980) J. Biol. Chem. b:6521-4.
  3. George, J. and Chirikjian, J.G. (1982) Proc. Natl. Acad. Sci. USA b:2432-6.
  4. Malyguine, E. et al. (1980) Gene 8:163-77.
  5. Clarke, C.M. and Hartley, B.S. (1979) Biochem. J. 177:49-62.
  6. Heininger, K. et al. (1977) Gene b:291-303.
  7. Makula, R.A. and Meagher, R.B. (1980) Nucl. Acids Res. 8:3125-31.
  8. Polisky, B. et al. (1975) Proc. Natl. Acad. Sci. USA 72:3310-4.
  9. Tikchonenko, T.I. et al. (1978) Gene 4:195-212.
  10. Hsu, M. and Berg, P. (1978) Biochemistry 17:131-8.
  11. Woodbury, C.P. et al. (1980) J. Biol. Chem. 255:11534-46.
  12. Halford, S.E. et al. (1986) Gene 41:173-81.
  13. Nasri, M. and Thomas, D. (1986) Nucl. Acids Res. 14:811-22.
  14. Gingeras, T.R. and Brooks, J.E. (1983) Proc. Natl. Acad. Sci. USA 80:402-6.
  15. Nasri, M. et al. (1985) FEBS Lett. 185:101-4.
  16. Nasri, M. and Thomas, D. (1987) Nucl. Acids Res. 15:7677-87.
  17. Pech, M. et al. (1979) Cell 18:883-93.
  18. Barany, F. (1988) Gene 65:149-65.
  19. Shinomiya, T. et al. (1982) J. Biochem. 92:1823-32.
  20. Kessler, C., unpublished observations.
  21. Grosskopt, R. and Kessler, C., unpublished observations.
  22. Takara Bio Inc., unpublished observations.

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