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

Abstract:

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

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Guo Y,Dong J,Zhou T,Auxillos J,Li T,Zhang W,Wang L,Shen Y,Luo Y,Zheng Y,Lin J,Chen GQ,Wu Q,Cai Y,Dai J

doi

10.1093/nar/gkv464

subject

Has Abstract

pub_date

2015-07-27 00:00:00

pages

e88

issue

13

eissn

0305-1048

issn

1362-4962

pii

gkv464

journal_volume

43

pub_type

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