Investigating Fungal Biosynthetic Pathways Using Heterologous Gene Expression: Fusarium sp. as a Heterologous Host

Mikkel Rank Nielsen; Jens Laurids Sørensen

doi:10.1007/978-1-0716-2273-5_4

Investigating Fungal Biosynthetic Pathways Using Heterologous Gene Expression: Fusarium sp. as a Heterologous Host

Mikkel Rank Nielsen^*, Jens Laurids Sørensen

^*Kontaktforfatter

Publikation: Bidrag til bog/antologi/rapport/konference proceeding › Bidrag til bog/antologi › Forskning › peer review

1 Citationer (Scopus)

Abstract

Heterologous expression of uncharacterized biosynthetic gene clusters is a popular strategy for exploring
the chemical potential of filamentous fungi. Here, we describe the process of PCR-amplifying fungal gene
clusters and re-assembling them in a cloning vector via target-associated recombination in Saccharomyces
cerevisiae. The gene cluster-carrying construct is validated and used to transform protoplasts of Fusarium
graminearum, a well-studied host that is able to express the gene cluster. Chemical analysis of transformants
expressing biosynthetic genes can lead to the detection and isolation of novel compounds, such as
polyketides.

Originalsprog	Engelsk
Titel	Engineering Natural Product Biosynthesis : Methods and Protocols
Redaktører	Elizabeth Skellam
Antal sider	22
Udgivelsessted	New York
Forlag	Springer
Publikationsdato	7 maj 2022
Sider	53-74
ISBN (Trykt)	978-1-0716-2272-8
ISBN (Elektronisk)	978-1-0716-2273-5
DOI	https://doi.org/10.1007/978-1-0716-2273-5_4
Status	Udgivet - 7 maj 2022

Navn	Methods in Molecular Biology
Vol/bind	2489
ISSN	1064-3745

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10.1007/978-1-0716-2273-5_4

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Citationsformater

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title = "Investigating Fungal Biosynthetic Pathways Using Heterologous Gene Expression: Fusarium sp. as a Heterologous Host",

abstract = "Heterologous expression of uncharacterized biosynthetic gene clusters is a popular strategy for exploringthe chemical potential of filamentous fungi. Here, we describe the process of PCR-amplifying fungal geneclusters and re-assembling them in a cloning vector via target-associated recombination in Saccharomycescerevisiae. The gene cluster-carrying construct is validated and used to transform protoplasts of Fusariumgraminearum, a well-studied host that is able to express the gene cluster. Chemical analysis of transformantsexpressing biosynthetic genes can lead to the detection and isolation of novel compounds, such aspolyketides.",

keywords = "Chromosomal integration, Fungi, Fusarium, Gene cluster, Heterologous expression, Homologous recombination, Protoplast, Protoplast-mediated transformation, TAR",

author = "Nielsen, {Mikkel Rank} and S{\o}rensen, {Jens Laurids}",

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Investigating Fungal Biosynthetic Pathways Using Heterologous Gene Expression: Fusarium sp. as a Heterologous Host. / Nielsen, Mikkel Rank; Sørensen, Jens Laurids.
Engineering Natural Product Biosynthesis: Methods and Protocols. red. / Elizabeth Skellam. New York: Springer, 2022. s. 53-74 (Methods in Molecular Biology, Bind 2489).

Publikation: Bidrag til bog/antologi/rapport/konference proceeding › Bidrag til bog/antologi › Forskning › peer review

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AU - Nielsen, Mikkel Rank

AU - Sørensen, Jens Laurids

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N2 - Heterologous expression of uncharacterized biosynthetic gene clusters is a popular strategy for exploringthe chemical potential of filamentous fungi. Here, we describe the process of PCR-amplifying fungal geneclusters and re-assembling them in a cloning vector via target-associated recombination in Saccharomycescerevisiae. The gene cluster-carrying construct is validated and used to transform protoplasts of Fusariumgraminearum, a well-studied host that is able to express the gene cluster. Chemical analysis of transformantsexpressing biosynthetic genes can lead to the detection and isolation of novel compounds, such aspolyketides.

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KW - Chromosomal integration

KW - Fungi

KW - Fusarium

KW - Gene cluster

KW - Heterologous expression

KW - Homologous recombination

KW - Protoplast

KW - Protoplast-mediated transformation

KW - TAR

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