Bidirectional histone-gene promoters in Aspergillus : characterization and application for multi-gene expression.

Autor: Rendsvig JKH; Department of Biotechnology and Biomedicine, Technical University of Denmark, 2800 Kongens Lyngby, Denmark., Workman CT; Department of Biotechnology and Biomedicine, Technical University of Denmark, 2800 Kongens Lyngby, Denmark., Hoof JB; Department of Biotechnology and Biomedicine, Technical University of Denmark, 2800 Kongens Lyngby, Denmark.
Jazyk: angličtina
Zdroj: Fungal biology and biotechnology [Fungal Biol Biotechnol] 2019 Dec 09; Vol. 6, pp. 24. Date of Electronic Publication: 2019 Dec 09 (Print Publication: 2019).
DOI: 10.1186/s40694-019-0088-3
Abstrakt: Background: Filamentous fungi are important producers of enzymes and bioactive secondary metabolites and are exploited for industrial purposes. Expression and characterization of biosynthetic pathways requires stable expression of multiple genes in the production host. Fungal promoters are indispensable for the accomplishment of this task, and libraries of promoters that show functionality across diverse fungal species facilitate synthetic biology approaches, pathway expression, and cell-factory construction.
Results: In this study, we characterized the intergenic region between the genes encoding histones H4.1 and H3, from five phylogenetically diverse species of Aspergillus , as bidirectional promoters (P h4h3 ). By expression of the genes encoding fluorescent proteins mRFP1 and mCitrine, we show at the translational and transcriptional level that this region from diverse species is applicable as strong and constitutive bidirectional promoters in Aspergillus nidulans . Bioinformatic analysis showed that the divergent gene orientation of h4.1 and h3 appears maintained among fungi, and that the P h4h3 display conserved DNA motifs among the investigated 85 Aspergilli. Two of the heterologous P h4h3 s were utilized for single-locus expression of four genes from the putative malformin producing pathway from Aspergillus brasiliensis in A. nidulans . Strikingly, heterologous expression of mlfA encoding the non-ribosomal peptide synthetase is sufficient for biosynthesis of malformins in A. nidulans , which indicates an iterative use of one adenylation domain in the enzyme. However, this resulted in highly stressed colonies, which was reverted to a healthy phenotype by co-expressing the residual four genes from the putative biosynthetic gene cluster.
Conclusions: Our study has documented that P h4h3 is a strong constitutive bidirectional promoter and a valuable new addition to the genetic toolbox of at least the genus Aspergillus .
Competing Interests: Competing interestsThe authors declare that they have no competing interests.
(© The Author(s) 2019.)
Databáze: MEDLINE
Externí odkaz: