【摘要】 通过生物信息学方法从耐辐射奇球菌 (D .radiodurans)全基因组数据库中查寻并克隆了编码过氧化氢酶(catalase ,Cat)的 16 11bp长CatB基因。将CatB基因连入 pKK2 2 3 3表达载体 ,转化Cat酶缺陷型大肠杆菌 (E .coliUM 2 )。转化菌裂解液PAGE酶活性染色分析表明 ,该基因表达产物具有Cat酶活性 ,电泳迁移位置与CatB位置相符。D .radioduransCatB基因的表达可使E .coliUM 2重建对低浓度H2 O2 的耐受力。更多还原

【Abstract】 A 1 611 bp length complete coding sequence of the catalase (Cat) was obtained through bioimformatical analysis of the database of D.radiodurans genome, and then was amplified from D. radiodurans genomic DNA by polymerase chain reaction. The amplified gene was cloned into pKK223 3 vector and transformed into E.coli UM2, a Cat deficient mutant. Staining of non denaturing polyacrylamide gels for Cat activity demonstrated that pKK223 3 insert encoded a Cat that co migrated with the CatB found in D.radiodurans cell lysates. Expression of Cat B gene from D.radiodurans in E.coli UM2 restored the resistance to H 2O 2 at low concentrations.更多还原