title: Expression and purification of active protein kinases from wheat germ extracts.
abstract: In vitro functional studies of eukaryotic kinases are often constrained by the availability of pure and -enzymatically active kinase of interest. Though numerous proteins have been synthesized by cell-based systems, in vivo production of properly folded, eukaryotic proteins remains a challenging task. Current wheat-germ-based cell-free in vitro translation systems present a plausible alternative for protein synthesis since majority of eukaryotic proteins could be obtained in their native folded form with general protocols. The use of special in vitro translation vectors with ligation-independent cloning sites and cleavable affinity tags eliminates further bottlenecks of the protein producing procedure and makes this system a reasonable method for simultaneous generation of active kinases.
authors.affiliation: Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Budapest, Hungary.
authors.forename: Boglárka
authors.initials: B
authors.lastname: Sonkoly
authors.forename: Viola
authors.initials: V
authors.lastname: Bardóczy
authors.forename: Tamás
authors.initials: T
authors.lastname: Mészáros
mesh_terms: D002474:Cell-Free System; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors / Q000235:genetics; D014176:Protein Biosynthesis; D011494:Protein Kinases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014908:Triticum / Q000235:genetics* / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D011494:Protein Kinases