title: Gene optimization mechanisms: a multi-gene study reveals a high success rate of full-length human proteins expressed in Escherichia coli.
abstract: The genetic code is universal, but recombinant protein expression in heterologous systems is often hampered by divergent codon usage. Here, we demonstrate that reprogramming by standardized multi-parameter gene optimization software and de novo gene synthesis is a suitable general strategy to improve heterologous protein expression. This study compares expression levels of 94 full-length human wt and sequence-optimized genes coding for pharmaceutically important proteins such as kinases and membrane proteins in E. coli. Fluorescence-based quantification revealed increased protein yields for 70% of in vivo expressed optimized genes compared to the wt DNA sequences and also resulted in increased amounts of protein that can be purified. The improvement in transgene expression correlated with higher mRNA levels in our analyzed examples. In all cases tested, expression levels using wt genes in tRNA-supplemented bacterial strains were outperformed by optimized genes expressed in non-supplemented host cells.
authors.affiliation: QIAGEN GmbH, QIAGEN Strasse 1, Hilden 40724, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Maertens
authors.forename: Anne
authors.initials: A
authors.lastname: Spriestersbach
authors.forename: Uritza
authors.initials: U
authors.lastname: von Groll
authors.forename: Udo
authors.initials: U
authors.lastname: Roth
authors.forename: Jan
authors.initials: J
authors.lastname: Kubicek
authors.forename: Michael
authors.initials: M
authors.lastname: Gerrits
authors.forename: Marcus
authors.initials: M
authors.lastname: Graf
authors.forename: Michael
authors.initials: M
authors.lastname: Liss
authors.forename: Daniela
authors.initials: D
authors.lastname: Daubert
authors.forename: Ralf
authors.initials: R
authors.lastname: Wagner
authors.forename: Frank
authors.initials: F
authors.lastname: Schäfer
mesh_terms: D003062:Codon / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D003062:Codon; D011506:Proteins; D011994:Recombinant Proteins