Directed evolution of ribosomal protein S1 for enhanced translational efficiency of high GC Rhodopseudomonas palustris DNA in Escherichia coli.
The expression of foreign DNA in Escherichia coli is important in biotechnological applications. However, the translation of genes from GC-rich organisms is inefficient in E. coli. To overcome this problem, we applied directed evolution to E. coli ribosomal protein S1. Two selected mutants enabled 12- and 8-fold higher expression levels from GC-rich DNA targets. General improvements in translation efficiency over a range of genes from Rhodopseudomonas palustris and E. coli was achieved using an S1 mutant selected against multiple genes from R. palustris. This method opens new opportunities for the expression of GC-rich genes in E. coli.
Bernstein
Jeffrey R
JR
Department of Chemical and Biomolecular Engineering, UCLA, Los Angeles, California 90095, USA.
Bulter
Thomas
T
Shen
Claire R
CR
Liao
James C
JC
D020121:5' Untranslated Regions / Q000235:genetics; D000595:Amino Acid Sequence; D001709:Biotechnology / Q000379:methods; D019020:Directed Molecular Evolution / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics*; D020862:GC Rich Sequence / Q000235:genetics; D007763:Lac Operon; D008969:Molecular Sequence Data; D016296:Mutagenesis; D014176:Protein Biosynthesis / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics; D012241:Rhodopseudomonas / Q000235:genetics*; D012269:Ribosomal Proteins / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D020121:5' Untranslated Regions; D012333:RNA, Messenger; D012269:Ribosomal Proteins; C015494:ribosomal protein S1; D001616:beta-Galactosidase