Codon Optimizing for Increased Membrane Protein Production: A Minimalist Approach
Reengineering a gene with synonymous codons is a popular approach for increasing production levels of recombinant proteins. Here we present a minimalist alternative to this method, which samples synonymous codons only at the second and third positions rather than the entire coding sequence. As demonstrated with two membrane-embedded transporters in Escherichia coli, the method was more effective than optimizing the entire coding sequence. The method we present is PCR based and requires three simple steps: (1) the design of two PCR primers, one of which is degenerate; (2) the amplification of a mini-library by PCR; and (3) screening for high-expressing clones.
Key wordsMembrane protein Protein expression Codon optimization Synonymous codon
This work was supported by a grant from the Swedish Research Council to DOD, and by the Novo Nordisk Foundation to MHHN.
Conflict of interest statement: The method considered here has been described in a patent submitted by CloneOpt AB (SE1451553-0). KM, ST, and DOD are founders and shareholders in CloneOpt AB.