A newly constructed Agrobacterium-mediated transformation system revealed the influence of nitrogen sources on the function of the LaeA regulator in Penicillium chrysogenum

Vu, Thang X.; Vu, Ha Hong; Nguyen, Giang T.; Vu, Hien; Dam, Linh Thi; Pham, Duc-Ngoc; Le, Diep Hong; Nguyen, Huy Quang; Tran, Van‐Tuan

doi:10.1016/j.funbio.2019.08.010

Cited by 8 publications

(4 citation statements)

References 65 publications

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“…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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“…However, the efficiency of AMT in particular strains is highly dependent on certain parameters, like the biological properties of the fungus, coculturing conditions, and the fungus‐to‐ Agrobacterium ratio, which need to be optimized accordingly [38, 64]. Additional examples of filamentous fungi in which this method has successfully applied include Fusarium proliferatum [65] , B. dothidea [62] , Penicillium chrysogenum [66] A. oryzae [67], and many others [24].…”

Section: Traditional Genetic Transformation Techniques In Filamentousmentioning

confidence: 99%

CRISPR/Cas9‐based genome engineering: A new breakthrough in the genetic manipulation of filamentous fungi

Ullah

Lin

Xie

et al. 2020

Biotech and App Biochem

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Filamentous fungi have several industrial, environmental, and medical applications. However, they are rarely utilized owing to the limited availability of full‐genome sequences and genetic manipulation tools. Since the recent discovery of the full‐genome sequences for certain industrially important filamentous fungi, CRISPR/Cas9 technology has drawn attention for the efficient development of engineered strains of filamentous fungi. CRISPR/Cas9 genome editing has been successfully applied to diverse filamentous fungi. In this review, we briefly discuss the use of common genetic transformation techniques as well as CRISPR/Cas9‐based systems in filamentous fungi. Furthermore, we describe potential limitations and challenges in the practical application of genome engineering of filamentous fungi. Finally, we provide suggestions and highlight future research prospects in the area.

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“…Gene knockout [15,16], gene complementation [17], and gene overexpression [16,18] are often used to identify the gene function of filamentous fungi, but selectable marker genes are required in the above experiments. If strains are sensitive to antibiotics or herbicides, which is helpful for screening target mutants.…”

Section: Introductionmentioning

confidence: 99%

Screening and Functional Verification of Selectable Marker Genes for Cordyceps militaris

Lou

Zhao

et al. 2021

Journal of Food Quality

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The selectable marker genes are necessary resistance genes for gene knockout, gene complementation, and gene overexpression in filamentous fungi. Moreover, the more sensitive the filamentous fungi are to antibiotics, the more helpful it is to screen the target transformants. In order to obtain the antibiotic (or herbicide) which can effectively inhibit the growth of Cordyceps militaris and verify the function of the corresponding resistance gene in C. militaris, the sensitivity of C. militaris to hygromycin and glufosinate ammonium was compared to determine the resistance gene that was more suitable for the screening of C. militaris transformants. The binary vector of the selectable marker gene was constructed by combining the double-joint PCR (DJ-PCR) method and the homologous recombination method, and the function of the selectable marker gene in C. militaris was verified by the Agrobacterium tumefaciens-mediated transformation method. The results showed that C. militaris was more sensitive to glufosinate ammonium than hygromycin. The growth of C. militaris could be completely inhibited by 250 μg/mL glufosinate ammonium. The expression cassette of the glufosinate ammonium resistance gene (bar gene) was successfully constructed by DJ-PCR. The binary vector pCAMBIA0390-Bar was successfully constructed by homologous recombination. The bar gene of the vector pCAMBIA0390-Bar was successfully integrated into the C. militaris genome and could be highly expressed in the transformants of C. militaris. This study will promote the identification of C. militaris gene function and reveal the biosynthetic pathways of bioactive components in C. militaris.

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A newly constructed Agrobacterium-mediated transformation system revealed the influence of nitrogen sources on the function of the LaeA regulator in Penicillium chrysogenum

Cited by 8 publications

References 65 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

CRISPR/Cas9‐based genome engineering: A new breakthrough in the genetic manipulation of filamentous fungi

Screening and Functional Verification of Selectable Marker Genes for Cordyceps militaris

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