IIS类限制性内切酶MboII的纯化及特性
Purification and properties of the MboII, a class-IIS restriction endonuclease.
作者信息
Sektas M, Kaczorowski T, Podhajska A J
机构信息
Department of Microbiology, University of Gdansk, Poland.
出版信息
Nucleic Acids Res. 1992 Feb 11;20(3):433-8. doi: 10.1093/nar/20.3.433.
After five purification steps a homogeneous preparation of endonuclease MboII was obtained, and several properties of the enzyme were determined. MboII is a monomer, with Mr under native and denaturing conditions being 47-49 x 10(3) Da. Endonuclease MboII is a basic protein (pI 8.3) which remains active when Mg2+ is replaced by Mn2+, Co2+, Ca2+, or Fe2+. MboII exhibits a star activity in the presence of some of the following reagents or ions: DMSO, glycerol, ethanol (and Co2+ or Mn2+ at pH 6). MboII does not bend DNA and is heat sensitive, losing activity after 15 min at 50 degrees C.
经过五步纯化后，获得了内切核酸酶MboII的均一制剂，并测定了该酶的几个特性。MboII是一种单体，在天然和变性条件下其相对分子质量为47 - 49×10³Da。内切核酸酶MboII是一种碱性蛋白质（pI 8.3），当Mg²⁺被Mn²⁺、Co²⁺、Ca²⁺或Fe²⁺取代时仍保持活性。MboII在存在下列某些试剂或离子时表现出星号活性：二甲基亚砜（DMSO）、甘油、乙醇（以及在pH 6时的Co²⁺或Mn²⁺）。MboII不会使DNA弯曲，并且对热敏感，在50℃下15分钟后失去活性。
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