【摘要】 这篇文章报道了金黄色葡萄球菌核酸酶A的克隆和在温控启动子P_RP_L调控下在E.coli中的高效表达。SDS-PAGE分析表明,核酸酶A的含量可占E.coli细胞总蛋白含量的60％。经有效的增溶和复性处理后,重组酶具有与天然酶相同的活力;N-末端氨基酸分析的结果指出,fMet在转译后被加工去除;对重组SNaseA在构象上的同一性也通过Phenyl-Su-perose疏水柱层析进行了分析。更多还原

【Abstract】 The staphylococcal nuclease A gene has been successfully cloned and overex-pressed in E. coli under the transcriptional control of the bacteriophage λ PRPL promoters regulated by the temperature sensitive repressers. The SDS-PAGE analysis demonstrates that the nuclease A is produced to the extent of as much as 60% of the total cellular protein . The N-terminal analysis of the nuclease A shows that the amino terminal formyl methionine residue of the enzyme is precisely processed. The recombinant nuclease A with full activity is finally obtained after appropriate solubilization-denaturation and renaturation. The conformational identity of the renatured SNaseA in different conditions is also studied by using Hydrophobic Interaction Chromatography on a phenyl-superose HR5/5 column.更多还原