ArticleWorld journal of microbiology & biotechnology2014
Screening of multi-copy mannanase recombinants of Pichia pastoris based on colony size.
Article in World journal of microbiology & biotechnology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- Heterologous Expression of Secreted Proteins from Phytophthora in Pichia pastoris Followed by Protein Purification by Immobilized Metal Ion Affinity (IMAC).Methods in molecular biology (Clifton, N.J.) · 2025Article
- Affinity-based and in a single step purification of recombinant horseradish peroxidase A2A isoenzyme produced by Pichia pastoris.Bioprocess and biosystems engineering · 2023Article
- Transcriptomic Analysis of the Influence of Methanol Assimilation on the Gene Expression in the RecombinantGenes · 2019Article
- Insights into the prevalence and underlying causes of clonal variation through transcriptomic analysis in Pichia pastoris.Applied microbiology and biotechnology · 2017Article
- Development of simple random mutagenesis protocol for the protein expression system in Pichia pastoris.Biotechnology for biofuels · 2016Article
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Authors and funding
5 authors.
Funding
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Abstract
Pichia pastoris pGAP (glyceraldehyde dehydrogenase promoter) expression system was widely used for the expression and production of heterologous proteins. Screening multi-copy recombinants was an effective strategy to improve the heterologous protein production in P. pastoris. Because multiple gene insertion events occurred with a low frequency, hundreds to thousands of antibiotic-resistance recombinants need to be screened. The common way of improving screening efficiency was to increase antibiotic concentration in screening plates. Here we developed a screening method by selecting small colonies from low-concentration antibiotic screening plates. This strategy greatly improved the probability of obtaining multi-copy mannanase gene (man) recombinants and it could replace the common strategy by increasing antibiotic concentration in screening plates. The further study in liquid shake flask cultures revealed that cell concentrations, growth rates and substrate consumption rates of recombinants gradually decreased with the increase in man copy number. This indicated that such a screening strategy was effective to screen multi-copy recombinants based on colony size.
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