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Fig. 2 | Microbial Cell Factories

Fig. 2

From: Refactoring the architecture of a polyketide gene cluster enhances docosahexaenoic acid production in Yarrowia lipolytica through improved expression and genetic stability

Fig. 2

Modular workflow for the genetic assembly of multigene clusters for genomic expression in Yarrowia lipolytica. The use of specific restriction enzymes enables the correct modular assembly of repetitive single gene modules into four-gene PUFA synthase gene clusters. First, single PUFA cluster genes are fused with a terminator. The resulting DNA fragments are integrated into a preconfigured promoter vector using Gibson assembly. The promoter-gene-terminator elements are excised from the vector using restriction digestion with AjuI (A) and subsequently integrated into an assembly vector in a defined order using unique restriction sites (B), which yields the complete cluster. The cluster is inserted into a shuttle vector between two homology domains using restriction and ligation. The cluster is then excised for chromosomal integration into Y. lipolytica (C). The figure was created using Biorender.com

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Microbial Cell Factories

ISSN: 1475-2859

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