Pichia pastoris Random Mutagenesis and Screening.

Christine Winkler, Joan Lin-Cereghino, Geoff P Lin-Cereghino

Abstract read

In one paragraph

Article in Methods in molecular biology (Clifton, N.J.), 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it

0cells of the map it votes in

0citing papers in PubMed

–field-weighted citation impact

1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

3 authors.

Christine WinklerInstitute of Molecular Biotechnology, Graz University of Technology, Graz, Austria.

Joan Lin-CereghinoDepartment of Biological Sciences, University of the Pacific, Stockton, CA, USA. jlincere@pacific.edu.

Geoff P Lin-CereghinoDepartment of Biological Sciences, University of the Pacific, Stockton, CA, USA. glincere@pacific.edu.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Random mutagenesis provides a valuable tool to elucidate gene function and to draw connections between gene products and molecular pathways. This chapter provides two protocols for the random mutagenesis of Komagataella phaffii, along with methods to screen for the resulting mutant phenotypes. Genes are first disrupted by the insertion of a non-homologous DNA cassette. In the next step, primers designed to anneal to the cassette enable the identification of the mutagenized locus. We applied our system in K. phaffii to enhance recombinant protein secretion of horseradish peroxidase (HRP) and β-galactosidase, respectively. The subsequent screening identified unexpected genes that were later confirmed to benefit recombinant protein secretion, validating the substantive potential of the methodology.

Indexed as

MutagenesisSaccharomycetalesbeta-GalactosidaseHorseradish PeroxidaseRecombinant Proteinsbeta-GalactosidaseHorseradish PeroxidaseRecombinant ProteinsGenome walkingInsertional mutagenesisKomagataella phaffiiPichia pastorisProtein secretionRestriction enzyme-mediated integration (REMI)

Identifiers

PMID41028479

DOI10.1016/j.femsyr.2004.06.016

What OpenQuestion holds

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.