Gene Cloning and Functional Analysis of the Luciferase from Luminous Syllids of the Genus Odontosyllis

Rie Yasuno; Yasuo Mitani; Yoshihiro Ohmiya

doi:10.1007/978-1-0716-2453-1_1

Gene Cloning and Functional Analysis of the Luciferase from Luminous Syllids of the Genus Odontosyllis

Methods Mol Biol. 2022:2524:3-15. doi: 10.1007/978-1-0716-2453-1_1.

Authors

Rie Yasuno¹, Yasuo Mitani², Yoshihiro Ohmiya^{3

4}

Affiliations

¹ Cellular and Molecular Biotechnology Research Institute, Advanced Industrial Science and Technology (AIST), Tsukuba, Japan.
² Bioproduction Research Institute, AIST, Sapporo, Japan. mitani-y@aist.go.jp.
³ Biomedical Research Institute, AIST, Osaka, Japan.
⁴ Osaka Institute of Technology (OIT), Osaka, Japan.

PMID: 35821459
DOI: 10.1007/978-1-0716-2453-1_1

Abstract

The marine fireworm Odontosyllis spp. produce the bluish-green bioluminescence (BL). Despite years of research, molecular mechanisms of this unique luciferin-luciferase reaction have not been elucidated. Recently, the genes encoding luciferases of O. undecimdonta and O. enopla have been identified. Here, we describe gene cloning techniques for the luciferase of Odontosyllis spp. from a small number of specimens using highly sensitive mass spectrometry analysis in combination with RNA-sequencing. The luciferase activities of the cloned cDNAs are confirmed by BL assay in vitro using recombinant protein expressed in mammalian cells.

Keywords: Bioluminescence (BL); Luciferase; Odontosyllis; RNA-sequencing; Recombinant expression.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cloning, Molecular
DNA, Complementary / genetics
DNA, Complementary / metabolism
Luciferases / metabolism
Mammals / genetics
Polychaeta*
Recombinant Proteins / metabolism

Substances

DNA, Complementary
Recombinant Proteins
Luciferases