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Volume 166, Issue 6

Multigenic engineering of the chloroplast genome in the green alga Open Access

Abstract

The chloroplast of microalgae such as represents an attractive chassis for light-driven production of novel recombinant proteins and metabolites. Methods for the introduction and expression of transgenes in the chloroplast genome (=plastome) of are well-established and over 100 different proteins have been successfully produced. However, in almost all reported cases the complexity of the genetic engineering is low, and typically involves introduction into the plastome of just a single transgene together with a selectable marker. In order to exploit fully the potential of the algal chassis it is necessary to establish methods for multigenic engineering in which many transgenes can be stably incorporated into the plastome. This would allow the synthesis of multi-subunit proteins and the introduction into the chloroplast of whole new metabolic pathways. In this short communication we report a proof-of-concept study involving both a combinatorial and serial approach, with the goal of synthesizing five different test proteins in the chloroplast. Analysis of the various transgenic lines confirmed the successful integration of the transgenes and accumulation of the gene products. However, the work also highlights an issue of genetic instability when using the same untranslated region for each of the transgenes. Our findings therefore help to define appropriate strategies for robust multigenic engineering of the algal chloroplast.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): Chlamydomonas , chloroplast , genetic engineering , microalgae and plastome
Funding
This study was supported by the:
  • Secretaría de Educación Superior, Ciencia, Tecnología e Innovación (Award n/a)
    • Principle Award Recipient: Marco Larrea-Alvarez
  • Biotechnology and Biological Sciences Research Council (Award BB/R016534/1)
    • Principle Award Recipient: Saul Purton
  • Biotechnology and Biological Sciences Research Council (Award BB/L002957/1)
    • Principle Award Recipient: Saul Purton
© 2020 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2020-04-06
2024-04-19
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Multigenic engineering of the chloroplast genome in the green alga Chlamydomonas reinhardtii
Microbiology 166, 510 (2020); https://doi.org/10.1099/mic.0.000910
/content/journal/micro/10.1099/mic.0.000910
/content/journal/micro/10.1099/mic.0.000910
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