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New destination vectors facilitate Modular Cloning for Chlamydomonas

Synthetic Biology is revolutionizing biological research by introducing principles of mechanical engineering, including the standardization of genetic parts and standardized part assembly routes. Both are realized in the Modular Cloning (MoClo) strategy. MoClo allows for the rapid and robust assembl...

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Autores principales: Niemeyer, Justus, Schroda, Michael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9279246/
https://www.ncbi.nlm.nih.gov/pubmed/35429260
http://dx.doi.org/10.1007/s00294-022-01239-x
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author Niemeyer, Justus
Schroda, Michael
author_facet Niemeyer, Justus
Schroda, Michael
author_sort Niemeyer, Justus
collection PubMed
description Synthetic Biology is revolutionizing biological research by introducing principles of mechanical engineering, including the standardization of genetic parts and standardized part assembly routes. Both are realized in the Modular Cloning (MoClo) strategy. MoClo allows for the rapid and robust assembly of individual genes and multigene clusters, enabling iterative cycles of gene design, construction, testing, and learning in short time. This is particularly true if generation times of target organisms are short, as is the case for the unicellular green alga Chlamydomonas reinhardtii. Testing a gene of interest in Chlamydomonas with MoClo requires two assembly steps, one for the gene of interest itself and another to combine it with a selection marker. To reduce this to a single assembly step, we constructed five new destination vectors. They contain genes conferring resistance to commonly used antibiotics in Chlamydomonas and a site for the direct assembly of basic genetic parts. The vectors employ red/white color selection and, therefore, do not require costly compounds like X-gal and IPTG. mCherry expression is used to demonstrate the functionality of these vectors.
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spelling pubmed-92792462022-07-15 New destination vectors facilitate Modular Cloning for Chlamydomonas Niemeyer, Justus Schroda, Michael Curr Genet Technical Note Synthetic Biology is revolutionizing biological research by introducing principles of mechanical engineering, including the standardization of genetic parts and standardized part assembly routes. Both are realized in the Modular Cloning (MoClo) strategy. MoClo allows for the rapid and robust assembly of individual genes and multigene clusters, enabling iterative cycles of gene design, construction, testing, and learning in short time. This is particularly true if generation times of target organisms are short, as is the case for the unicellular green alga Chlamydomonas reinhardtii. Testing a gene of interest in Chlamydomonas with MoClo requires two assembly steps, one for the gene of interest itself and another to combine it with a selection marker. To reduce this to a single assembly step, we constructed five new destination vectors. They contain genes conferring resistance to commonly used antibiotics in Chlamydomonas and a site for the direct assembly of basic genetic parts. The vectors employ red/white color selection and, therefore, do not require costly compounds like X-gal and IPTG. mCherry expression is used to demonstrate the functionality of these vectors. Springer Berlin Heidelberg 2022-04-16 2022 /pmc/articles/PMC9279246/ /pubmed/35429260 http://dx.doi.org/10.1007/s00294-022-01239-x Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Technical Note
Niemeyer, Justus
Schroda, Michael
New destination vectors facilitate Modular Cloning for Chlamydomonas
title New destination vectors facilitate Modular Cloning for Chlamydomonas
title_full New destination vectors facilitate Modular Cloning for Chlamydomonas
title_fullStr New destination vectors facilitate Modular Cloning for Chlamydomonas
title_full_unstemmed New destination vectors facilitate Modular Cloning for Chlamydomonas
title_short New destination vectors facilitate Modular Cloning for Chlamydomonas
title_sort new destination vectors facilitate modular cloning for chlamydomonas
topic Technical Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9279246/
https://www.ncbi.nlm.nih.gov/pubmed/35429260
http://dx.doi.org/10.1007/s00294-022-01239-x
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