A paradigm shift is underway within the methodology of heterologous protein expression. Specifically, researchers are moving away from conventional techniques of cloning genes from cDNA libraries and moving toward the rational design and de novo synthesis of entire protein-coding sequences from pre-annealed oligonucleotides (Libertini and Di Donato, 1992; Gustafsson et al, 2004). It was the invention of polymerase chain reaction (PCR) that allowed efficient construction of synthetic genes. Since then, the steadily increasing accuracy and decreasing cost of oligonucleotide synthesis (now as low as $0.10 per base; Carlson, 2003; Carr et al, 2004; Kong et al, 2007, see Figure 1) has created a research environment in which gene synthesis offers three main advantages over molecular cloning: cost efficiency, scope and flexibility of redesign (Libertini and Di Donato, 1992). As a result, the emerging field of synthetic biology is highly motivated to improve this approach, as it seeks to expand the sophistication of human-engineered genetic architectures, leading ultimately to the synthesis of entire genomes (Yount et al, 2000; Smith et al, 2003).
Molecular Systems Biology
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