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Gene. 2018 Jan 15;640:14-20. doi: 10.1016/j.gene.2017.10.012. Epub 2017 Oct 7.

Isolation and characterization of a stress-responsive gene encoding a CHRD domain-containing protein from a halotolerant green alga.

Author information

1
Applied Environmental Biology Laboratory, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
2
Applied Environmental Biology Laboratory, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: hmhide@phs.osaka-u.ac.jp.
3
The Kansai Electric Power Co., Inc., Advanced Technology Laboratory, Keihanna Engineering Center, 1-7 Seika-cho, Souraku-gun, Kyoto 619-0237, Japan.
4
Pharmainformatics and Pharmacometrics Laboratory, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
5
International Center for Biotechnology, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
6
Department of Applied Life Science, Sojo University, 4-22-1 Ikeda, Nishiku, Kumamoto 860-0082, Japan.

Abstract

The genetic basis of stress resistance in extremophilic microalgae is not well studied. In this study, a gene of unknown function, the cluster58 or CL58 gene, was identified from the halotolerant green alga Chlamydomonas W80 and characterized. The CL58 gene encodes a protein containing a domain of unknown function, the CHRD domain, and a putative secretory signaling sequence at its N-terminus. The levels of CL58 mRNA increased in response to high copper levels and low temperatures. When the CL58 gene was heterologously expressed as a fusion gene with the NanoLuc luciferase gene in Chlamydomonas reinhardtii, a majority of the NanoLuc activity was detected in the culture medium compared with that in the intracellular fraction. A mutagenic analysis revealed that the putative secretory signaling sequence was sufficient for the secretion of the CL58-NanoLuc fusion protein. In addition, we expressed the protein encoded by the CL58 gene in Escherichia coli; the recombinant, soluble protein was then purified. In summary, we identified a novel gene from C. W80 that appears to encode a stress-responsive, CHRD domain-containing secreted protein.

KEYWORDS:

Chlamydomonas reinhardtii; Recombinant protein; Secretory protein; Stress response; Unknown function

PMID:
29017964
DOI:
10.1016/j.gene.2017.10.012
[Indexed for MEDLINE]

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