1. Development of a chloroplast expression system for Dunaliella salina.
- Author
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Chen HH, Zheng QX, Yu F, Xie SR, and Jiang JG
- Subjects
- Promoter Regions, Genetic, Plants, Genetically Modified genetics, Plants, Genetically Modified metabolism, Genetic Engineering methods, Microalgae genetics, Microalgae metabolism, Chlorophyceae genetics, Chlorophyceae metabolism, Chlorophyta genetics, Chlorophyta metabolism, Homologous Recombination, Gene Expression, Chloroplasts genetics, Chloroplasts metabolism, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, Genetic Vectors metabolism, Transformation, Genetic
- Abstract
Dunaliella salina is an innovative expression system due to its distinct advantages such as high salt tolerance, low susceptibility to contamination, and the absence of the cell wall. While nuclear transformation has been extensively studied, research on D. salina chloroplast transformation remains in the preliminary stages. In this study, we established an efficient chloroplast expression system for D. salina using Golden Gate assembly. We developed a D. salina toolkit comprising essential components such as chloroplast-specific promoters, terminators, homologous fragments, and various vectors. We confirmed its functionality by expressing the EGFP protein. Moreover, we detailed the methodology of the entire construction process. This expression system enables the specific targeting of foreign genes through simple homologous recombination, resulting in stable expression in chloroplasts. The toolkit achieved a relatively high transformation efficiency within a shorter experimental cycle. Consequently, the construction and utilization of this toolkit have the potential to enhance the efficiency of transgenic engineering in D. salina and advance the development of microalgal biofactories., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)
- Published
- 2024
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