ArticleScientific reports2016
Non-canonical integration events in Pichia pastoris encountered during standard transformation analysed with genome sequencing.
Article in Scientific reports, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed.
- Omics-Based Expression Cassette for Heterologous Protein Production inACS synthetic biology · 2026Article
- Pichia pastoris Genomic Resources: Accessibility, Recommendations, and Pitfalls.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Establishing Reference Genes for Pichia pastoris Quantitative Real-Time Polymerase Chain Reaction Analyses.Methods in molecular biology (Clifton, N.J.) · 2026Article
- A toolkit for facilitating markerless integration of expression cassettes in Komagataella phaffii via CRISPR/Cas9.Microbial cell factories · 2025Article
- Omics-driven onboarding of the carotenoid producing red yeast Xanthophyllomyces dendrorhous CBS 6938.Applied microbiology and biotechnology · 2024Article
- A supernumerary synthetic chromosome in Komagataella phaffii as a repository for extraneous genetic material.Microbial cell factories · 2023Article
- Use of an on/off tetracycline riboswitch to control protein production in Komagataella phaffii.AMB Express · 2023Article
- Domain Truncation in Hispidin Synthase Orthologs from Non-Bioluminescent Fungi Does Not Lead to Hispidin Biosynthesis.International journal of molecular sciences · 2023Article
- PFrontiers in bioengineering and biotechnology · 2023Article
- Whole Genome Sequencing Analysis of Effects of CRISPR/Cas9 inJournal of fungi (Basel, Switzerland) · 2022Article
- The fluorescent protein iLOV as a reporter for screening of high-yield production of antimicrobial peptides in Pichia pastoris.Microbial biotechnology · 2022Article
- Competent Cell Preparation and Transformation of Pichia pastoris.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Screening and evaluation of the strong endogenous promoters in Pichia pastoris.Microbial cell factories · 2021Article
- Improvements of the productivity and saccharification efficiency of the cellulolytic β-glucosidase D2-BGL in Pichia pastoris via directed evolution.Biotechnology for biofuels · 2021Article
- Engineered yeast genomes accurately assembled from pure and mixed samples.Nature communications · 2021Article
- Exchange of endogenous and heterogeneous yeast terminators in Pichia pastoris to tune mRNA stability and gene expression.Nucleic acids research · 2020Article
- Rationale-based selection of optimal operating strategies and gene dosage impact on recombinant protein production in Komagataella phaffii (Pichia pastoris).Microbial biotechnology · 2020Article
- Orthologous promoters from related methylotrophic yeasts surpass expression of endogenous promoters of Pichia pastoris.AMB Express · 2020Article
- Review
- Process development for the continuous production of heterologous proteins by the industrial yeast, Komagataella phaffii.Biotechnology and bioengineering · 2018Article
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The non-conventional yeast Pichia pastoris is a popular host for recombinant protein production in scientific research and industry. Typically, the expression cassette is integrated into the genome via homologous recombination. Due to unknown integration events, a large clonal variability is often encountered consisting of clones with different productivities as well as aberrant morphological or growth characteristics. In this study, we analysed several clones with abnormal colony morphology and discovered unpredicted integration events via whole genome sequencing. These include (i) the relocation of the locus targeted for replacement to another chromosome (ii) co-integration of DNA from the E. coli plasmid host and (iii) the disruption of untargeted genes affecting colony morphology. Most of these events have not been reported so far in literature and present challenges for genetic engineering approaches in this yeast. Especially, the presence and independent activity of E. coli DNA elements in P. pastoris is of concern. In our study, we provide a deeper insight into these events and their potential origins. Steps preventing or reducing the risk for these phenomena are proposed and will help scientists working on genetic engineering of P. pastoris or similar non-conventional yeast to better understand and control clonal variability.
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