Busso Didier, Kim Rosalind, Kim Sung-Hou
Department of Chemistry, University of California, Berkeley, 314 Calvin Lab MS5230, CA 94720-5230, USA.
J Biochem Biophys Methods. 2003 Mar 28;55(3):233-40. doi: 10.1016/s0165-022x(03)00049-6.
For structural and functional genomics programs, new high-throughput methods to obtain well-expressing and highly soluble proteins are essential. Here, we describe a rapid procedure to express recombinant proteins in an Escherichia coli cell-free system using a 96-well format. The identification of soluble proteins is performed by the Dot Blot procedure using an anti-His tag antibody. The applications and the automation of this method are described.
对于结构和功能基因组学计划而言,获得表达良好且高度可溶蛋白质的新型高通量方法至关重要。在此,我们描述了一种使用96孔板形式在大肠杆菌无细胞系统中表达重组蛋白的快速程序。通过使用抗His标签抗体的斑点印迹程序来鉴定可溶性蛋白质。本文还描述了该方法的应用及自动化操作。