Start your expression with an optimal DNA sequence using GeneOptimizer
One of the advantages of synthetic oligonucleotide sequences is the opportunity to make changes to the nucleotide sequence without altering the amino acid sequence.
Codon usage is different in each organism, so it can be challenging to get expression in heterologous expression systems. Furthermore, regulatory sequence elements at the transcriptional, translational, and mRNA levels can limit recombinant protein expression. With our gene synthesis and DNA fragments services, we offer free sequence optimization using our proprietary GeneOptimizer software. During the optimization process your DNA sequence will be adjusted to represent your defined host codon usage, and expression-limiting sequence motifs will be eliminated to provide you with an optimal sequence for reliable protein expression.
Our sequence optimization can help increase protein expression levels compared to wild type expression. Figure 4 illustrates the results of an internal study comparing expression of wild type sequences with sequence-optimized counterparts generated with GeneOptimizer software, for five randomly selected different proteins and antibodies. Protein levels of the sequence-optimized constructs are clearly increased compared to the corresponding wild type constructs. Further examples of the impact of sequence optimization on protein expression can also be found in a large-scale expression study published by Fath et al. .
Gene synthesis, subcloning into Invitrogen pcDNA3.3 vector, and plasmid preparation were performed by our scientists in Regensburg, Germany. Intracellular proteins (RFP, CREB1, OPRM1) were expressed in adherent HEK293 cells (2 mL scale) in triplicate and quantified by α-His western blot analysis. Antibodies (VRC-01, 10E8) were expressed in Invitrogen FreeStyle 293 cells (30 mL scale) in duplicate, and expression levels were determined via the BLItz system (ForteBio). The mean protein expression levels for each of the five proteins, for wild type and optimized sequences, are graphed in Figure 4. Protein expression was normalized to expression of the wild type sequence.
Increased protein yield is not the only advantage of sequence optimization. Sequences that have been traditionally difficult to clone due to the presence of unstable elements can be subjected to sequence optimization, which can result in improved cloning efficiencies and higher expression levels (2). Additionally, using GeneArt optimized sequences can help with overall experimental cost savings (see the Cost Effectiveness tab at the top of this page).
If your project doesn’t require the use of wild type sequences, let us help by starting with an optimized DNA sequence.
- Fath S, Bauer AP et al. (2011) Multiparameter RNA and Codon Optimization: A Standardized Tool to Assess and Enhance Autologous Mammalian Gene Expression. PLoS One 6(3): e17596.