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An inducible expression system in the alga Nannochloropsis gaditana controlled by the nitrate reductase promoter

An inducible expression system in the alga Nannochloropsis gaditana controlled by the nitrate reductase promoter
An inducible expression system in the alga Nannochloropsis gaditana controlled by the nitrate reductase promoter

The marine microalgal species Nannochloropsis gaditana has emerged as a model organism for research into algal-derived biofuels due to its ability to produce large amounts of lipids. In addition, this species has biotechnological potential owing to its ease of transformation and availability of a sequenced and annotated genome. The ability to control expression of transgenes in an inducible manner is an important requirement for many genetic engineering approaches and as such, inducible promoters are an important component of the molecular toolkit for a host organism. We designed an expression vector containing a ~ 1.3 kb region upstream of the endogenous nitrate reductase gene of N. gaditana and demonstrated this is capable of promoting the expression of a heterologous enhanced green fluorescent protein (eGFP) reporter. In the presence of ammonium, expression of the eGFP reporter was undetectable; however, in the presence of nitrate, eGFP expression is induced. In addition, we demonstrated how altering the nitrogen source of the growth media can be used to precisely control expression. We have shown that the nitrate reductase promoter can be used as a powerful molecular tool for heterologous protein expression in N. gaditana, further contributing to the development of this species as a candidate strain for biotechnology.

Biofuels, Heterologous expression, Inducible promoter, Nannochloropsis, Nitrate reductase, Synthetic biology
0921-8971
269–279
Jackson, Harry O.
984d0348-9117-4cd1-b729-942447300f27
Berepiki, Adokiye
1be9b7f8-011f-454c-bc9e-244b2c07799a
Baylay, Alison J.
08ea69f8-9910-4c8b-86a9-602e45b8a44e
Terry, Matthew J.
a8c2cd6b-8d35-4053-8d77-3841c2427c3b
Moore, C. Mark
7ec80b7b-bedc-4dd5-8924-0f5d01927b12
Bibby, Thomas S.
e04ea079-dd90-4ead-9840-00882de27ebd
Jackson, Harry O.
984d0348-9117-4cd1-b729-942447300f27
Berepiki, Adokiye
1be9b7f8-011f-454c-bc9e-244b2c07799a
Baylay, Alison J.
08ea69f8-9910-4c8b-86a9-602e45b8a44e
Terry, Matthew J.
a8c2cd6b-8d35-4053-8d77-3841c2427c3b
Moore, C. Mark
7ec80b7b-bedc-4dd5-8924-0f5d01927b12
Bibby, Thomas S.
e04ea079-dd90-4ead-9840-00882de27ebd

Jackson, Harry O., Berepiki, Adokiye, Baylay, Alison J., Terry, Matthew J., Moore, C. Mark and Bibby, Thomas S. (2019) An inducible expression system in the alga Nannochloropsis gaditana controlled by the nitrate reductase promoter. Journal of Applied Phycology, 31, 269–279. (doi:10.1007/s10811-018-1510-6).

Record type: Article

Abstract

The marine microalgal species Nannochloropsis gaditana has emerged as a model organism for research into algal-derived biofuels due to its ability to produce large amounts of lipids. In addition, this species has biotechnological potential owing to its ease of transformation and availability of a sequenced and annotated genome. The ability to control expression of transgenes in an inducible manner is an important requirement for many genetic engineering approaches and as such, inducible promoters are an important component of the molecular toolkit for a host organism. We designed an expression vector containing a ~ 1.3 kb region upstream of the endogenous nitrate reductase gene of N. gaditana and demonstrated this is capable of promoting the expression of a heterologous enhanced green fluorescent protein (eGFP) reporter. In the presence of ammonium, expression of the eGFP reporter was undetectable; however, in the presence of nitrate, eGFP expression is induced. In addition, we demonstrated how altering the nitrogen source of the growth media can be used to precisely control expression. We have shown that the nitrate reductase promoter can be used as a powerful molecular tool for heterologous protein expression in N. gaditana, further contributing to the development of this species as a candidate strain for biotechnology.

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10.1007%2Fs10811-018-1510-6 - Version of Record
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Accepted/In Press date: 9 May 2018
e-pub ahead of print date: 25 May 2018
Published date: February 2019
Keywords: Biofuels, Heterologous expression, Inducible promoter, Nannochloropsis, Nitrate reductase, Synthetic biology

Identifiers

Local EPrints ID: 421354
URI: http://eprints.soton.ac.uk/id/eprint/421354
ISSN: 0921-8971
PURE UUID: 2d127e01-16bf-4482-b1a6-05830709a503
ORCID for Matthew J. Terry: ORCID iD orcid.org/0000-0001-5002-2708

Catalogue record

Date deposited: 05 Jun 2018 16:30
Last modified: 20 Jul 2019 01:17

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Contributors

Author: Harry O. Jackson
Author: Adokiye Berepiki
Author: C. Mark Moore
Author: Thomas S. Bibby

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