YeastFab: the design and construction of standard biological parts for metabolic engineering in Saccharomyces cerevisiae.

摘要: It is a routine task in metabolic engineering to introduce multicomponent pathways into heterologous host for production of metabolites. However, this process sometimes may take weeks months due the lack standardized genetic tools. Here, we present method design and construction biological parts based on native genes regulatory elements Saccharomyces cerevisiae. We have developed highly efficient protocols (termed YeastFab Assembly) synthesize these as parts, which can be used assemble transcriptional units single-tube reaction. In addition, characterization assays are using reporter constructs calibrate function promoters. Furthermore, assembled transcription either assayed individually or applied construct multi-gene pathways, targets genomic locus receiving plasmid effectively, through simple vitro Finally, β-carotene biosynthesis pathway an example, demonstrate that our allows us not only test several days, but also optimize combinatorial assembly hundreds parts.