Work

Strong Translation Elongation Factor 1-Alpha (tef1α) Promoter Reporter Systems for Aspergillus niger

Year: 2025

Type: article

Abstract: High Resolution Image Download MS PowerPoint Slide Strong promoters are crucial for microbial metabolic engineering and synthetic biology, enabling high-level gene expression and pathway optimization. In this study, we have identified the translation elongation factor 1-α gene promoter ( tef1 p) with its first intron ( tef1IN p) as the strongest promoter reported to date in Aspergillus niger . It exhibited more than 20-fold higher activity in a methylumbelliferone reporter assay than the commonly used glyceraldehyde-3-phosphate dehydrogenase ( gpdA ) promoter. The t ef1 INp demonstrated consistent performance across multiple reporter genes, including EGFP, mCherry, mRFP1, and GUS, yielding strong fluorescence and staining signals. Its robustness was further validated in a coexpression system using P2A peptides, which enabled efficient bicistronic expression with a streamlined vector design. Additionally, we achieved precise nuclear localization and colocalization using H2B- and H3-based fusion constructs with EGFP and mRFP1. Using this system, we successfully identified the nuclear localization signals (NLS) of H2B and H3 in A. niger . Overall, this work establishes versatile and high-performance genetic tools for advanced applications in fungal synthetic biology.

Source: ACS Synthetic Biology

DOI: 10.1021/acssynbio.5c00368