title: Expression of soluble recombinant proteins in a cell-free system using a 96-well format.
abstract: 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.
authors.affiliation: Department of Chemistry, University of California, Berkeley, 314 Calvin Lab MS5230, CA 94720-5230, USA. djbusso@lbl.gov
authors.forename: Didier
authors.initials: D
authors.lastname: Busso
authors.forename: Rosalind
authors.initials: R
authors.lastname: Kim
authors.forename: Sung-Hou
authors.initials: SH
authors.lastname: Kim
mesh_terms: D002474:Cell-Free System / Q000378:metabolism*; D004867:Equipment Design; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods; D015202:Protein Engineering / Q000295:instrumentation / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D012371:Robotics / Q000295:instrumentation* / Q000379:methods; D012995:Solubility
chemical_list: D011994:Recombinant Proteins