title: Coordinated induction of multi-gene pathways in Saccharomyces cerevisiae.
abstract: Bacterial operons are nature's tool for regulating and coordinating multi-gene expression in prokaryotes. They are also a gene architecture commonly used in the biosynthesis of many pharmaceutically important compounds and industrially useful chemicals. Despite being an important eukaryotic production host, Saccharomyces cerevisiae has never had such gene architecture. Here, we report the development of a system to assemble and regulate a multi-gene pathway in S. cerevisiae. Full pathways can be constructed using pre-made parts from a plasmid toolbox. Subsequently, through the use of a yeast strain containing a stably integrated gene switch, the assembled pathway can be regulated using a readily available and inexpensive compound-estradiol-with extremely high sensitivity (10 nM). To demonstrate the use of the system, we assembled the five-gene zeaxanthin biosynthetic pathway in a single step and showed the ligand-dependent coordinated expression of all five genes as well as the tightly regulated production of zeaxanthin. Compared with a previously reported constitutive zeaxanthin pathway, our inducible pathway was shown to have 50-fold higher production level.
authors.affiliation: Department of Chemical and Biomolecular Engineering and Departments of Chemistry, Biochemistry, and Bioengineering, Center for Biophysics and Computational Biology, Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Liang
authors.forename: Jonathan C
authors.initials: JC
authors.lastname: Ning
authors.forename: Huimin
authors.initials: H
authors.lastname: Zhao
mesh_terms: D004958:Estradiol / Q000494:pharmacology; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005818:Genetic Engineering / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D024341:Xanthophylls / Q000096:biosynthesis; D065146:Zeaxanthins
chemical_list: D012333:RNA, Messenger; D024341:Xanthophylls; D065146:Zeaxanthins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004958:Estradiol