title: Continuous-exchange protein-synthesizing systems.
abstract: Protein synthesis in cell-free systems is an emerging technology already competing with in vivo expression methods. In this chapter the basic principles of continuous-exchange protein synthesizing systems, and protocols for Escherichia coli and wheat germ translation and transcription-translation systems are described. The ways to improve substrate supply in cell-free systems and mRNA design for eukaryotic system are discussed. Correct folding of the synthesized protein is demonstrated and discussed in detail.
authors.affiliation: Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow, Russia.
authors.forename: Vladimir A
authors.initials: VA
authors.lastname: Shirokov
authors.forename: Aigar
authors.initials: A
authors.lastname: Kommer
authors.forename: Vyacheslav A
authors.initials: VA
authors.lastname: Kolb
authors.forename: Alexander S
authors.initials: AS
authors.lastname: Spirin
mesh_terms: D002474:Cell-Free System; D004926:Escherichia coli / Q000378:metabolism; D008967:Molecular Biology / Q000379:methods*; D014176:Protein Biosynthesis; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry* / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D014158:Transcription, Genetic; D014908:Triticum / Q000378:metabolism
chemical_list: D011506:Proteins; D012333:RNA, Messenger