ArticleFungal biology and biotechnology2023
Evaluation of reference genes for transcript analyses in Komagataella phaffii (Pichia pastoris).
Article in Fungal biology and biotechnology, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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8 citing papers in PubMed, 16 citations in OpenAlex.
- Engineering genetic elements for microbial protein expression systems: Advances, challenges, applications, and prospects.Synthetic and systems biotechnology · 2026Review
- Comparative and functional genomics of yeast Pichia kudriavzevii RWT identify potential genes involved in aflatoxin B1 detoxification.Molecular genetics and genomics : MGG · 2026Article
- Sterol regulation in Komagataella phaffii: Identification of sterol regulatory element binding proteins and their activation pathway.The Journal of biological chemistry · 2025Article
- Higher induction temperatures and the native secretion signal peptide promote rye prolamin 75k γ-secalin production in Komagataella phaffii.Microbial cell factories · 2025Article
- Structural Homology Fails to Predict Secretion Efficiency inInternational journal of molecular sciences · 2025Article
- Article
- Chances and drawbacks of derepressed recombinant enzyme production in continuous cultivations withFrontiers in bioengineering and biotechnology · 2025Article
- Bi-directionalized promoter systems allow methanol-free production of hard-to-express peroxygenases with Komagataella Phaffii.Microbial cell factories · 2024Article
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7 authors at 1 institution in 1 country.
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Abstract
backgroundThe yeast Komagataella phaffii (Pichia pastoris) is routinely used for heterologous protein expression and is suggested as a model organism for yeast. Despite its importance and application potential, no reference gene for transcript analysis via RT-qPCR assays has been evaluated to date. In this study, we searched publicly available RNASeq data for stably expressed genes to find potential reference genes for relative transcript analysis by RT-qPCR in K. phaffii. To evaluate the applicability of these genes, we used a diverse set of samples from three different strains and a broad range of cultivation conditions. The transcript levels of 9 genes were measured and compared using commonly applied bioinformatic tools.
resultsWe could demonstrate that the often-used reference gene ACT1 is not very stably expressed and could identify two genes with outstandingly low transcript level fluctuations. Consequently, we suggest the two genes, RSC1, and TAF10 to be simultaneously used as reference genes in transcript analyses by RT-qPCR in K. phaffii in future RT-qPCR assays.
conclusionThe usage of ACT1 as a reference gene in RT-qPCR analysis might lead to distorted results due to the instability of its transcript levels. In this study, we evaluated the transcript levels of several genes and found RSC1 and TAF10 to be extremely stable. Using these genes holds the promise for reliable RT-qPCR results.
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