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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Bibliographic Details
Published in:Current genetics 2022-08, Vol.68 (3-4), p.531-536
Main Authors: Niemeyer, Justus, Schroda, Michael
Format: Article
Language:English
Subjects:
Antibiotics
Aquatic plants
Assembly
Biochemistry
Biological research
Biomedical and Life Sciences
Cell Biology
Chlamydomonas
Cloning
Cloning vectors
Cycle protein
Expression vectors
Genes
Life Sciences
Mechanical engineering
Microbial Genetics and Genomics
Microbiology
Plant Sciences
Proteomics
Standardization
Technical Note
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Summary: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.
ISSN:0172-8083
1432-0983
1432-0983
DOI:10.1007/s00294-022-01239-x