Novel endogenous promoters for genetic engineering of the marine microalga Nannochloropsis gaditana CCMP526

Ramarajan, M; Fabris, M; Abbriano, RM; Pernice, M; Ralph, PJ

Novel endogenous promoters for genetic engineering of the marine microalga Nannochloropsis gaditana CCMP526

Ramarajan, M Fabris, M

Abbriano, RM

Pernice, M

Ralph, PJ

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Publication Type:: Journal Article
Citation:: Algal Research, 2019, 44
Issue Date:: 2019-12-01

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Field	Value	Language
dc.contributor.author	Ramarajan, M	en_US
dc.contributor.author	Fabris, M https://orcid.org/0000-0003-2910-2089	en_US
dc.contributor.author	Abbriano, RM https://orcid.org/0000-0002-5754-5461	en_US
dc.contributor.author	Pernice, M https://orcid.org/0000-0002-3431-2104	en_US
dc.contributor.author	Ralph, PJ https://orcid.org/0000-0002-3103-7346	en_US
dc.date.issued	2019-12-01	en_US
dc.identifier.citation	Algal Research, 2019, 44	en_US
dc.identifier.issn	2211-9264	en_US
dc.identifier.uri	http://hdl.handle.net/10453/137350
dc.description.abstract	© 2019 Elsevier B.V. Nannochloropsis is a marine microalga from the Eustigmatophyceae stramenopile lineage that has been studied extensively due to a broad range of industrial applications, mostly related to their oil and pigment production. However, tools to genetically engineer members of this group, and therefore further understand and maximise their industrial potential are still limited. In order to expand the potential industrial uses of this organism, several molecular tools, including gene promoters of different strength, are needed. A comprehensive and diverse set of well-characterized promoters is key to a number of genetic engineering and synthetic biology applications, such as the assembly of complex biological functions or entire metabolic pathways. In this study, we measured the promoter activity of three endogenous constitutive promoters from N. gaditana genes EPPSII (Nga02101); HSP90 (Nga00934); ATPase (Nga06354.1) in driving the expression of a Sh ble- mVenus fluorescent reporter fusion protein. Through a combined approach that includes flow cytometry, RT-qPCR and immunoblotting, we profiled the activity of these promoters at both the transcript and protein level. Two promoters HSP90 (Nga00934) and EPPSII (Nga02101) outperformed the widely used β-tubulin promoter, exhibiting 4.5 and 3.1-fold higher mVenus fluorescence, respectively. A third promoter ATPase (Nga06354.1) was also able to drive the expression of transgenes, albeit at lower levels. We show that the new promoters identified in this study are valuable tools, which can be used for genetic engineering and functional genetics studies in N. gaditana.	en_US
dc.relation.ispartof	Algal Research	en_US
dc.relation.isbasedon	10.1016/j.algal.2019.101708	en_US
dc.rights	info:eu-repo/semantics/embargoedAccess
dc.title	Novel endogenous promoters for genetic engineering of the marine microalga Nannochloropsis gaditana CCMP526	en_US
dc.type	Journal Article
utslib.citation.volume	44	en_US
utslib.for	0607 Plant Biology	en_US
utslib.for	0604 Genetics	en_US
utslib.for	0904 Chemical Engineering	en_US
utslib.for	1003 Industrial Biotechnology	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - C3 - Climate Change Cluster
utslib.copyright.status	open_access	*
utslib.copyright.embargo	2021-12-02T00:00:00+1100
pubs.publication-status	Published	en_US
pubs.volume	44	en_US

Abstract:

© 2019 Elsevier B.V. Nannochloropsis is a marine microalga from the Eustigmatophyceae stramenopile lineage that has been studied extensively due to a broad range of industrial applications, mostly related to their oil and pigment production. However, tools to genetically engineer members of this group, and therefore further understand and maximise their industrial potential are still limited. In order to expand the potential industrial uses of this organism, several molecular tools, including gene promoters of different strength, are needed. A comprehensive and diverse set of well-characterized promoters is key to a number of genetic engineering and synthetic biology applications, such as the assembly of complex biological functions or entire metabolic pathways. In this study, we measured the promoter activity of three endogenous constitutive promoters from N. gaditana genes EPPSII (Nga02101); HSP90 (Nga00934); ATPase (Nga06354.1) in driving the expression of a Sh ble- mVenus fluorescent reporter fusion protein. Through a combined approach that includes flow cytometry, RT-qPCR and immunoblotting, we profiled the activity of these promoters at both the transcript and protein level. Two promoters HSP90 (Nga00934) and EPPSII (Nga02101) outperformed the widely used β-tubulin promoter, exhibiting 4.5 and 3.1-fold higher mVenus fluorescence, respectively. A third promoter ATPase (Nga06354.1) was also able to drive the expression of transgenes, albeit at lower levels. We show that the new promoters identified in this study are valuable tools, which can be used for genetic engineering and functional genetics studies in N. gaditana.

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http://hdl.handle.net/10453/137350