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A wide-range integrative yeast expression vector system based on Arxula adeninivorans-derived elements.

Yaroslav Terentiev¹, Almudena Huarto Pico, Erik Böer

¹Institut für Pflanzengenetik und Kulturpflanzenforschung, Corrensstrasse 3, 06466 Gatersleben, Germany.

Journal of Industrial Microbiology & Biotechnology

|June 4, 2004

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A novel integration vector successfully integrated into diverse yeast species genomes. This enables high-level heterologous gene expression across various yeast hosts for biotechnological applications.

Area of Science:

Microbiology
Molecular Biology
Biotechnology

Background:

Developing efficient tools for genetic manipulation in diverse yeast species is crucial for expanding their biotechnological potential.
Existing integration vectors may have limited applicability across different yeast genera.

Purpose of the Study:

To develop and test a novel integration vector for heterologous gene expression in alternative yeast species.
To assess the efficiency of genome integration and reporter gene expression in various yeast hosts.

Main Methods:

An Arxula adeninivorans integration vector containing a 25S rDNA targeting sequence, a TEF1 promoter, and a hygromycin B resistance gene (hph) was constructed.
The vector was introduced into Saccharomyces cerevisiae, Debaryomyces hansenii, Debaryomyces polymorphus, Hansenula polymorpha, and Pichia pastoris.

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Heterologous gene expression was evaluated using a green fluorescent protein (GFP) reporter gene.

Main Results:

The integration vector successfully integrated into the genomes of all tested yeast species.
Recombinant strains across all species demonstrated high-level heterologous gene expression.
The green fluorescent protein (GFP) reporter gene confirmed successful and functional gene expression.

Conclusions:

The developed integration vector is effective for genome integration and high-level heterologous gene expression in a broad range of yeast species.
This tool expands the genetic engineering toolkit for diverse yeast hosts, facilitating advancements in synthetic biology and biotechnology.