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Red Fluorescent Protein Variant with a Dual-Peak Emission of Fluorescence

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Abstract

The marine environment is a rich reservoir of diverse biological entities, many of which possess unique properties that are of immense value to biotechnological applications. One such example is the red fluorescent protein derived from the coral Discosoma sp. This protein, encoded by the DsRed gene, has been the subject of extensive research due to its potential applications in various fields. In the study, a variant of the red fluorescent protein was generated through random mutagenesis using the DsRed2 gene as a template. The process employed error-prone PCR (epPCR) to introduce random mutations, leading to the isolation of twelve gene variants. Among these, one variant stood out due to its unique spectral properties, exhibiting dual fluorescence emission at both 480 nm (green) and 550 nm (red). This novel variant was expressed in both Escherichia coli and zebrafish (Danio rerio) muscle, confirming the dual fluorescence emission in both model systems. One of the immediate applications of this novel protein variant is in ornamental aquaculture. The dual fluorescence can serve as a unique marker or trait, enhancing the aesthetic appeal of aquatic species in ornamental settings.

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The data and material used in this study are available upon request from the authors.

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Acknowledgements

We would like to express our gratitude to the Molecular Biology Laboratory at the Federal University of Rio Grande (FURG) and the Molecular Microbiology and Proteins Laboratory (LaMMP) at the Federal University of Rio de Janeiro (UFRJ) for their assistance, resources, and expertise, which were instrumental in the successful completion of our research.

Funding

This study was funded by CNPq (MCTI/CNPQ Call No. 454689/2014–4) and by CAPES, which provided the master’s scholarship. L.F. Marins is a research fellow from CNPq (Proc. no. 307304/2022-1).

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Authors and Affiliations

  1. Departamento de Biología, Universidad del Valle (UV), Cali, Valle del Cauca, Colombia

    Natalia Ossa-Hernández

  2. Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Universidade Federal do Rio Grande (FURG), Rio Grande, RS, Brazil

    Luis Fernando Marins

  3. Departamento de Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ, Brazil

    Rodrigo Volcan Almeida

  4. Departamento de Fisiologia e Farmacologia, Instituto de Biologia, Universidade Federal de Pelotas (UFPEL), Campus Universitário Capão do Leão s/n, CEP, Pelotas, RS, 96160-000, Brazil

    Daniela Volcan Almeida

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  1. Natalia Ossa-Hernández
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  2. Luis Fernando Marins
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  3. Rodrigo Volcan Almeida
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection, writing of the main manuscript text, and analysis were performed by N. O.-H. and D.V. A. All authors reviewed the manuscript.

Corresponding authors

Correspondence to Natalia Ossa-Hernández or Daniela Volcan Almeida.

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All experimental procedures were conducted in accordance with the Ethics for Animal Use Committee from the Federal University of Rio Grande, where this study was carried out (Protocol Pq024/2015). All participants gave their consent to participate.

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Ossa-Hernández, N., Marins, L.F., Almeida, R.V. et al. Red Fluorescent Protein Variant with a Dual-Peak Emission of Fluorescence. Mar Biotechnol 25, 1099–1109 (2023). https://doi.org/10.1007/s10126-023-10262-z

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