Efficient gene targeting in Penicillium chrysogenum using novel Agrobacterium-mediated transformation approaches

Boer, Paulo de; Bronkhof, Jurian; Dukiќ, Karolina; Kerkman, Richard; Touw, Hesselien; Berg, Marco van den; Offringa, Remko

doi:10.1016/j.fgb.2013.08.012

Cited by 13 publications

(6 citation statements)

References 30 publications

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“…Sun et al [ 310 ] reported the use of Agrobacterium tumefaciens -mediated transformation (ATMT). de Boer et al [ 311 ] used also an ATMT approach to investigate gene disruption efficiency (see below). Recently, Vu et al [ 312 ] developed an improved ATMT system for P. chrysogenum with two different selection markers, which yielded 5009 ± 96 transformants per 10 6 spores.…”

Section: A Growing Toolkitmentioning

confidence: 99%

Penicillium chrysogenum, a Vintage Model with a Cutting-Edge Profile in Biotechnology

et al. 2022

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The discovery of penicillin entailed a decisive breakthrough in medicine. No other medical advance has ever had the same impact in the clinical practise. The fungus Penicillium chrysogenum (reclassified as P. rubens) has been used for industrial production of penicillin ever since the forties of the past century; industrial biotechnology developed hand in hand with it, and currently P. chrysogenum is a thoroughly studied model for secondary metabolite production and regulation. In addition to its role as penicillin producer, recent synthetic biology advances have put P. chrysogenum on the path to become a cell factory for the production of metabolites with biotechnological interest. In this review, we tell the history of P. chrysogenum, from the discovery of penicillin and the first isolation of strains with high production capacity to the most recent research advances with the fungus. We will describe how classical strain improvement programs achieved the goal of increasing production and how the development of different molecular tools allowed further improvements. The discovery of the penicillin gene cluster, the origin of the penicillin genes, the regulation of penicillin production, and a compilation of other P. chrysogenum secondary metabolites will also be covered and updated in this work.

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Section: A Growing Toolkitmentioning

confidence: 99%

Penicillium chrysogenum, a Vintage Model with a Cutting-Edge Profile in Biotechnology

et al. 2022

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“…Such an approach has the potential to allow At MT delivery of separate partial selection cassettes, in a similar approach to that used very successfully in the bipartite or split-marker approach to enhancing targeted gene disruption in fungi [ 219 ]. Future implementation may further enhance gene targeting abilities in fungi [ 237 , 238 ].…”

Section: The Future and Further Maximizing The Impact Of Atmentioning

confidence: 99%

A silver bullet in a golden age of functional genomics: the impact of Agrobacterium-mediated transformation of fungi

Idnurm

Bailey

Cairns

et al. 2017

Fungal Biol Biotechnol

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The implementation of Agrobacterium tumefaciens as a transformation tool revolutionized approaches to discover and understand gene functions in a large number of fungal species. A. tumefaciens mediated transformation (AtMT) is one of the most transformative technologies for research on fungi developed in the last 20 years, a development arguably only surpassed by the impact of genomics. AtMT has been widely applied in forward genetics, whereby generation of strain libraries using random T-DNA insertional mutagenesis, combined with phenotypic screening, has enabled the genetic basis of many processes to be elucidated. Alternatively, AtMT has been fundamental for reverse genetics, where mutant isolates are generated with targeted gene deletions or disruptions, enabling gene functional roles to be determined. When combined with concomitant advances in genomics, both forward and reverse approaches using AtMT have enabled complex fungal phenotypes to be dissected at the molecular and genetic level. Additionally, in several cases AtMT has paved the way for the development of new species to act as models for specific areas of fungal biology, particularly in plant pathogenic ascomycetes and in a number of basidiomycete species. Despite its impact, the implementation of AtMT has been uneven in the fungi. This review provides insight into the dynamics of expansion of new research tools into a large research community and across multiple organisms. As such, AtMT in the fungi, beyond the demonstrated and continuing power for gene discovery and as a facile transformation tool, provides a model to understand how other technologies that are just being pioneered, e.g. CRISPR/Cas, may play roles in fungi and other eukaryotic species.

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“…Cả hai chủng P. chrysogenum đều bị ức chế hoàn toàn bởi kháng sinh nourseothricin ở nồng độ 50 µg/ml và nồng độ này thấp hơn rất nhiều so với nồng độ 500 µg/ml ở công bố trước đây cho loài P. chrysogenum [10]. Đối với kháng sinh phleomycin, sinh trưởng của hai chủng bị ức chế hoàn toàn ở nồng độ 150 µg/ml (Hình 4A).…”

Section: Mức độ Mẫn Cảm Kháng Sinh Và Bước đầU Chuyển Gen Vào Nấm Sợiunclassified

“…Như vậy, chúng tôi đã thành công trong việc chuyển gen vào nấm P. chrysogenum sử dụng vi khuẩn A. tumefaciens và marker kháng kháng sinh nourseothricin. Nồng độ nourseothricin sử dụng trong nghiên cứu này là 50 µg/ml thấp hơn 10 lần so với công bố của De-Boer và cs (2013) [10].…”

Section: Mức độ Mẫn Cảm Kháng Sinh Và Bước đầU Chuyển Gen Vào Nấm Sợiunclassified

Identification of biological characteristics and prelimilary research on genetic transformation of the filamentous fungus Penicillium chrysogenum originated from Vietnam

Tạo¹,

Tuấn²

2017

NST

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Penicillium chrysogenum is a well-known filamentous fungus for production of penicillin and some valuable secondary metabolites. In this study, we indicated that two strains VTCC-F1170 and VTCC-F1172 identified as P. chrysogenum, which are preserved at Vietnam Type Culture Collection (VTCC) of Vietnam National University Hanoi, exhibited the ability of antibiotic production to inhibit the tested bacterium Staphylococcus aureus on the agar plates by diffusion assays. Additional analyses of the morphological characteristics and the rDNA ITS (internal transcribed spacer) sequence confirmed that the identification of both strains as P. chrysogenum was completely accurate. These important evidences guaranteed that the fungal strains are reliable for the researches on genetic engineering of P. chrysogenum. Experimental assays of antibiotic susceptibility showed that the growth of both strains VTCC-F1170 and VTCC-F1172 was completely inhibited by nourseothricin at 50 μg/ml and phleomycin at 150 μg/ml. Using the Agrobacterium tumefaciens-mediated transformation method, we have succesfully transferred the nourseothricin resistance marker into the genome of the VTCC-F1170 strain.

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Efficient gene targeting in Penicillium chrysogenum using novel Agrobacterium-mediated transformation approaches

Cited by 13 publications

References 30 publications

Penicillium chrysogenum, a Vintage Model with a Cutting-Edge Profile in Biotechnology

Penicillium chrysogenum, a Vintage Model with a Cutting-Edge Profile in Biotechnology

A silver bullet in a golden age of functional genomics: the impact of Agrobacterium-mediated transformation of fungi

Identification of biological characteristics and prelimilary research on genetic transformation of the filamentous fungus Penicillium chrysogenum originated from Vietnam

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