Gene optimization

If you want to obtain maximum expression yields, it is a good idea to adapt the gene at question to the target host. There is a number of parameters which have an impact on the expression yield.

Today, gene synthesis is a very efficient and affordable process which allows to tweak a given gene sequence strategically in order to improve expression yields. By applying rational design principles to a gene sequence underlying a target protein, money and time can be saved for the downstream process.

Parameters which affect expression yields, are:

GC content
Codon bias (preference of a particular subset of codons for a given expression host)
mRNA destabilizing motifs
mRNA secondary structure
secondary ORFs (extended reading frames in the second or third frame or on the opposing strand)

Taking all these parameters (and other constraints such as specific restriction sites) into account simultaneously is a time consuming and difficult process. Therefore, I suggest that you use suitable design software which automates the process, such as Leto.

Or, even better, outsource the process altogether. For instance, Entelechon provides the optimization of a gene as part of the fee-for-service gene synthesis.