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. Author manuscript; available in PMC: 2015 Feb 9.
Published in final edited form as: Methods Mol Biol. 2013;1073:85–106. doi: 10.1007/978-1-62703-625-2_9

Figure 1.

Figure 1

(a) Preparation of each gene expression cassette using OE-PCR. Promoters (Px, Px+1), genes (Gx, Gx+1), and terminators (Tx, Tx+1) are individually PCR-amplified and joined together by OE-PCR. The resulting two cassettes are fused through the in vivo homologous recombination (HR) process. To generate an overlap of approximately 50 bp, the reverse primer used to amplify Tx contains a sequence of the first 20-25 nucleotides of Px+1, and the forward primer used to amplify Px+1 contains a sequence of the last 20-25 nucleotides of Tx. (b) One-step method for assembly of a biochemical pathway using in vivo homologous recombination (HR) in S. cerevisiae.