SibEnzyme restriction enzymes database

Список ферментов

Bls I

Enzyme name Bls I
Prototype BlsI
SKU SE-E533
Turbo version Not available
High-concentration version Not available
Recognition site Неверный формат сайта узнавания
Source Bacillus simplex 23
Optimal buffer SE-buffer W
Optimal temperature 30 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
1010501007550
Unit definition One unit is defined as the amount of enzyme required to hydrolyze at least one of three canonical sites 5`-G(5mC)NG(5mC)-3`/3`-CGN(5mC)G-5` in 1 μg of linearized plasmid pFsp4HI3 in 1 hour at 30°C in a total reaction volume of 50 μl. As a result a linearized plasmid pFsp4HI3 disappears (see run 4 in the figure). BlsI activity assay on DNA pFsp4HI3/DriI Lanes: 1 and 7 - 1 Kb SE DNA-markers. 2 - Control DNA pFsp4HI3/DriI, 3 - 0.5 μl Bls I (1/5) 4 - 1 μl Bls I (1/5) 5 - 2 μl Bls I (1/5) 6 - 1 μl of undiluted BlsI 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 three canonical sites 5`-G(5mC)NG(5mC)-3`/3`-CGN(5mC)G-5` [2].
Storage conditions 10 mM Tris-HCl (pH 7.5); 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 5 units of enzyme for 16 hours at 30°C in a total reaction volume of 50 μl.
Methylation sensitivity The enzyme cleaves C5-methylated DNA and doesn't cut unmodified DNA [1].
Supplied with enzyme 10 X SE-buffer W
Notes
References
  1. Chernukhin V.A., Tomilova J.E., Chmuzh E.V., Sokolova O.O., Dedkov V.S., Degtyarev S.Kh. Bacterial strain Bacillus simplex - producer of BlsI site specific endonuclease. // Russian Federation patent RU 2322494 C1 (2006). Chmuzh E.V., Kashirina Yu.G., Tomilova Yu.E., Chernukhin V.A., Okhapkina S.S., Gonchar D. A., Dedkov V.S., Abdurashitov M. A., Degtyarev S. Kh. Gene cloning, comparative analysis of the protein structures from Fsp4HI restriction-modification system and biochemical characterization of the recombinant DNA methyltransferase M.Fsp4HI. // Molecular Biology, V.41, No 1, p. 43-50 (2007)

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Bls I

Enzyme name Bls I
Prototype BlsI
SKU SE-E533
Turbo version Not available
High-concentration version Not available
Recognition site Неверный формат сайта узнавания
Source Bacillus simplex 23
Optimal buffer SE-buffer W
Optimal temperature 30 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
1010501007550
Unit definition One unit is defined as the amount of enzyme required to hydrolyze at least one of three canonical sites 5`-G(5mC)NG(5mC)-3`/3`-CGN(5mC)G-5` in 1 μg of linearized plasmid pFsp4HI3 in 1 hour at 30°C in a total reaction volume of 50 μl. As a result a linearized plasmid pFsp4HI3 disappears (see run 4 in the figure). BlsI activity assay on DNA pFsp4HI3/DriI Lanes: 1 and 7 - 1 Kb SE DNA-markers. 2 - Control DNA pFsp4HI3/DriI, 3 - 0.5 μl Bls I (1/5) 4 - 1 μl Bls I (1/5) 5 - 2 μl Bls I (1/5) 6 - 1 μl of undiluted BlsI 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 three canonical sites 5`-G(5mC)NG(5mC)-3`/3`-CGN(5mC)G-5` [2].
Storage conditions 10 mM Tris-HCl (pH 7.5); 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 5 units of enzyme for 16 hours at 30°C in a total reaction volume of 50 μl.
Methylation sensitivity The enzyme cleaves C5-methylated DNA and doesn't cut unmodified DNA [1].
Supplied with enzyme 10 X SE-buffer W
Notes
References
  1. Chernukhin V.A., Tomilova J.E., Chmuzh E.V., Sokolova O.O., Dedkov V.S., Degtyarev S.Kh. Bacterial strain Bacillus simplex - producer of BlsI site specific endonuclease. // Russian Federation patent RU 2322494 C1 (2006). Chmuzh E.V., Kashirina Yu.G., Tomilova Yu.E., Chernukhin V.A., Okhapkina S.S., Gonchar D. A., Dedkov V.S., Abdurashitov M. A., Degtyarev S. Kh. Gene cloning, comparative analysis of the protein structures from Fsp4HI restriction-modification system and biochemical characterization of the recombinant DNA methyltransferase M.Fsp4HI. // Molecular Biology, V.41, No 1, p. 43-50 (2007)