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

^*Corresponding author for this work

Research output: Contribution to book/anthology/report/conference proceeding › Book chapter › Research › peer-review

1 Citation (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.

Original language	English
Title of host publication	Engineering Natural Product Biosynthesis : Methods and Protocols
Editors	Elizabeth Skellam
Number of pages	22
Place of Publication	New York
Publisher	Springer
Publication date	7 May 2022
Pages	53-74
ISBN (Print)	978-1-0716-2272-8
ISBN (Electronic)	978-1-0716-2273-5
DOIs	https://doi.org/10.1007/978-1-0716-2273-5_4
Publication status	Published - 7 May 2022

Series	Methods in Molecular Biology
Volume	2489
ISSN	1064-3745

Keywords

Chromosomal integration
Fungi
Fusarium
Gene cluster
Heterologous expression
Homologous recombination
Protoplast
Protoplast-mediated transformation
TAR

Access to Document

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

AUB Link

Search for the material in Aalborg University Library's search engine

Cite this

@inbook{68ecc48ec7bf4022bb0095dfa100a6dc,

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}",

year = "2022",

month = may,

day = "7",

doi = "10.1007/978-1-0716-2273-5_4",

language = "English",

isbn = "978-1-0716-2272-8",

series = "Methods in Molecular Biology",

publisher = "Springer",

pages = "53--74",

editor = "Elizabeth Skellam",

booktitle = "Engineering Natural Product Biosynthesis",

address = "Germany",

}

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. ed. / Elizabeth Skellam. New York: Springer, 2022. p. 53-74 (Methods in Molecular Biology, Vol. 2489).

Research output: Contribution to book/anthology/report/conference proceeding › Book chapter › Research › peer-review

TY - CHAP

T1 - Investigating Fungal Biosynthetic Pathways Using Heterologous Gene Expression: Fusarium sp. as a Heterologous Host

AU - Nielsen, Mikkel Rank

AU - Sørensen, Jens Laurids

PY - 2022/5/7

Y1 - 2022/5/7

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.

AB - 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.

KW - Chromosomal integration

KW - Fungi

KW - Fusarium

KW - Gene cluster

KW - Heterologous expression

KW - Homologous recombination

KW - Protoplast

KW - Protoplast-mediated transformation

KW - TAR

UR - https://link.springer.com/book/10.1007/978-1-0716-2273-5

UR - http://www.scopus.com/inward/record.url?scp=85130000441&partnerID=8YFLogxK

U2 - 10.1007/978-1-0716-2273-5_4

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

M3 - Book chapter

SN - 978-1-0716-2272-8

T3 - Methods in Molecular Biology

SP - 53

EP - 74

BT - Engineering Natural Product Biosynthesis

A2 - Skellam, Elizabeth

PB - Springer

CY - New York

ER -

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

Abstract

Keywords

Access to Document

AUB Link

Other files and links

Cite this