title: Critical reflections on synthetic gene design for recombinant protein expression.
abstract: Gene synthesis enables the exploitation of the degeneracy of the genetic code to boost expression of recombinant protein targets for structural studies. This has created new opportunities to obtain structural information on proteins that are normally present in low abundance. Unfortunately, synthetic gene expression occasionally leads to insoluble or misfolded proteins. This could be remedied by recent insights in the effect of codon usage on translation initiation and elongation. In this review, we discuss the interplay between optimal gene and vector design to enhance expression in a particular host and highlight the benefits and potential pitfalls associated with protein expression from synthetic genes.
authors.affiliation: European Molecular Biology Laboratory, Hamburg Unit, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Annabel Ha
authors.initials: AH
authors.lastname: Parret
authors.affiliation: Protein Expression and Purification Core Facility, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg, Germany.
authors.forename: Hüseyin
authors.initials: H
authors.lastname: Besir
authors.affiliation: European Molecular Biology Laboratory, Hamburg Unit, Notkestrasse 85, 22607 Hamburg, Germany. Electronic address: r.meijers@embl-hamburg.de.
authors.forename: Rob
authors.initials: R
authors.lastname: Meijers
mesh_terms: D003062:Codon / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D017510:Protein Folding; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*
chemical_list: D003062:Codon; D012333:RNA, Messenger; D011994:Recombinant Proteins