Lee C C, Wong D W, Robertson G H
Western Regional Research Center, USDA-ARS, Albany, California 94710, USA.
J Protein Chem. 2001 Apr;20(3):233-7. doi: 10.1023/a:1010904109747.
Libraries of modified genes are often screened during the process of genetically engineering enzymes with specifically tailored activities. It is important, therefore, to create expression systems which allow for the rapid screening of many clones. We developed an Escherichia coli expression system which will secrete enzymes into the growth medium. We describe the first reported expression of barley alpha-amylase in E. coli. The enzyme is secreted onto solid media containing starch to produce easily visualized halos. In addition, the enzyme is secreted into liquid media in an intact, active form.
在对酶进行基因工程改造以获得特定定制活性的过程中,常常需要筛选修饰基因文库。因此,创建能够快速筛选众多克隆的表达系统非常重要。我们开发了一种大肠杆菌表达系统,该系统可将酶分泌到生长培养基中。我们描述了大麦α-淀粉酶在大肠杆菌中的首次报道表达。该酶被分泌到含有淀粉的固体培养基上,产生易于观察到的晕圈。此外,该酶以完整、有活性的形式分泌到液体培养基中。