Figure 4.

DNA assembler method developed by Zhao and coworkers [61•]. Multiple gene expression cassettes (encoding promoter-gene-terminator) are prepared by splicing PCR. Each fragment contains sequence homology to adjacent fragments at its termini enabling crossover events at both ends. All fragments are co-transformed and correctly assembled in S. cerevisiae with a linearized vector (or a helper fragment, not shown in the figure), yielding a plasmid encoding a functional pathway (or integration of a functional pathway into yeast chromosome). This method should significantly simplify the protein engineering of biochemical pathways and has various potential applications including biofuels production.