SibEnzyme restriction enzymes database

Список ферментов

Glu I

Enzyme name Glu I
Prototype GluI
SKU SE-E519
Turbo version Not available
High-concentration version Not available
Recognition site
5'… G(5mC)▼NG(5mC) …3'
3'… (5mC)GN▲(5mC)G …5'
Source Glacial ice bacterium GL24
Optimal buffer SE-buffer Y
Optimal temperature 37 °C
Inactivation temperature 80 °C
Buffer activity
BGOWYROSE
75752550100100
Unit definition One unit is defined as the amount of enzyme required to hydrolyze completely a unique site 5`-G(5mC)NG(5mC)-3`/3`-(5mC)GN(5mC)G-5` in 1 μg of linearized plasmid pFsp4HI3/DriI in 1 hour at 37°C in a total reaction volume of 50 μl. As a result a linearized plasmid pFsp4HI3 disappears (see run 4 in the figure). GluI digestion of recognition sequences with three 5-methylcytosines results in additional bands appearance (runs 4-6 in the figure). GluI activity assay on DNA pFsp4HI3/DriI Lanes: 1 and 6 - 1 Kb SE DNA-markers. 2 - Control DNA pFsp4HI3/DriI, 3 - 0.5 μl Glu I 4 - 1 μl Glu I 5 - 2 μl Glu I Products were separated in 1,4% agarose gel in Buffer TAE.
Assayed on DNA pFsp4HI3/DriI is a linearized plasmid pFsp4HI3, which carries a gene of DNA-methyltransferase M.Fsp4HI and includes a unique GluI site:5`-G(5mC)NG(5mC)-3`/3`-(5mC)GN(5mC)G-5` [2].
Storage conditions 10 mM KH 2 PO 4 (pH 7.45); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA, 50% glycerol; Store at -20°C.
Ligation
Nonspecific hydrolysis No detectable degradation of 1μg of Lambda DNA was observed after incubation with 1 units of enzyme for 16 hours at 37°C in a total reaction volume of 50 μl.
Methylation sensitivity The enzyme cleaves C5-methylated DNAand does not cut unmodified DNA [1].
Supplied with enzyme 10 X SE-buffer Y
Notes When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
References
  1. Chernukhin V.A., Chmuzh E.V., Tomilova J.E., Nayakshina T.N., Dedkov V.S., Degtyarev S.Kh. Bacterial strain Glacial ice bacterium - producer of GluI site specific endonuclease. // Russian Federation patent RU 2322492 C1 (2006). Zhilkina O.A., Repin V.E., Degtyarev S.Kh. Preparation method of restriction endonuclease SfaNI, that recognizes and cleaves DNA sequence 5'-GCATC(N)5^, 3'-CGTAG(N)9^.// Soviet Union patent SU 1442546 (1988). (In Russian)
Application
  1. V.A. Chernukhin, E.V. Chmuzh, Yu.E. Tomilova, T.N. Nayakshina, D.A. Gonchar, V.S. Dedkov, S.Kh. Degtyarev A novel site-specific endonuclease GluI recognizes methylated DNA sequence 5’-G(5mC)^NG(5mC)-3’/3’-(5mC)GN^(5mC)G. // Translated from "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 2, pp 13-17, 2007 Application: Chernukhin V.A, Abdurashitov M.A., Tomilov V.N., Gonchar D.A., Degtyarev S.Kh. Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro and in silico // Translated from "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 4, pp 19-27, 2007 Kravets AP, Mousseau TA, Litvinchuk AV, Ostermiller Sh, Vengen GS, Grodzinski DM. Changes in wheat DNA methylation pattern after chronic seed gamma-irradiation. // Tsitol Genet. 2010 Sep-Oct;44(5):18-22. Russian.

Поиск по названию

Glu I

Enzyme name Glu I
Prototype GluI
SKU SE-E519
Turbo version Not available
High-concentration version Not available
Recognition site
5'… G(5mC)▼NG(5mC) …3'
3'… (5mC)GN▲(5mC)G …5'
Source Glacial ice bacterium GL24
Optimal buffer SE-buffer Y
Optimal temperature 37 °C
Inactivation temperature 80 °C
Buffer activity
BGOWYROSE
75752550100100
Unit definition One unit is defined as the amount of enzyme required to hydrolyze completely a unique site 5`-G(5mC)NG(5mC)-3`/3`-(5mC)GN(5mC)G-5` in 1 μg of linearized plasmid pFsp4HI3/DriI in 1 hour at 37°C in a total reaction volume of 50 μl. As a result a linearized plasmid pFsp4HI3 disappears (see run 4 in the figure). GluI digestion of recognition sequences with three 5-methylcytosines results in additional bands appearance (runs 4-6 in the figure). GluI activity assay on DNA pFsp4HI3/DriI Lanes: 1 and 6 - 1 Kb SE DNA-markers. 2 - Control DNA pFsp4HI3/DriI, 3 - 0.5 μl Glu I 4 - 1 μl Glu I 5 - 2 μl Glu I Products were separated in 1,4% agarose gel in Buffer TAE.
Assayed on DNA pFsp4HI3/DriI is a linearized plasmid pFsp4HI3, which carries a gene of DNA-methyltransferase M.Fsp4HI and includes a unique GluI site:5`-G(5mC)NG(5mC)-3`/3`-(5mC)GN(5mC)G-5` [2].
Storage conditions 10 mM KH 2 PO 4 (pH 7.45); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA, 50% glycerol; Store at -20°C.
Ligation
Nonspecific hydrolysis No detectable degradation of 1μg of Lambda DNA was observed after incubation with 1 units of enzyme for 16 hours at 37°C in a total reaction volume of 50 μl.
Methylation sensitivity The enzyme cleaves C5-methylated DNAand does not cut unmodified DNA [1].
Supplied with enzyme 10 X SE-buffer Y
Notes When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
References
  1. Chernukhin V.A., Chmuzh E.V., Tomilova J.E., Nayakshina T.N., Dedkov V.S., Degtyarev S.Kh. Bacterial strain Glacial ice bacterium - producer of GluI site specific endonuclease. // Russian Federation patent RU 2322492 C1 (2006). Zhilkina O.A., Repin V.E., Degtyarev S.Kh. Preparation method of restriction endonuclease SfaNI, that recognizes and cleaves DNA sequence 5'-GCATC(N)5^, 3'-CGTAG(N)9^.// Soviet Union patent SU 1442546 (1988). (In Russian)
Application
  1. V.A. Chernukhin, E.V. Chmuzh, Yu.E. Tomilova, T.N. Nayakshina, D.A. Gonchar, V.S. Dedkov, S.Kh. Degtyarev A novel site-specific endonuclease GluI recognizes methylated DNA sequence 5’-G(5mC)^NG(5mC)-3’/3’-(5mC)GN^(5mC)G. // Translated from "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 2, pp 13-17, 2007 Application: Chernukhin V.A, Abdurashitov M.A., Tomilov V.N., Gonchar D.A., Degtyarev S.Kh. Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro and in silico // Translated from "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 4, pp 19-27, 2007 Kravets AP, Mousseau TA, Litvinchuk AV, Ostermiller Sh, Vengen GS, Grodzinski DM. Changes in wheat DNA methylation pattern after chronic seed gamma-irradiation. // Tsitol Genet. 2010 Sep-Oct;44(5):18-22. Russian.