Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit
| Autor: | Alejandra A. Schiavon Osorio, Baojun Wang, Alistair J. McCormick, Anton Puzorjov, Grant A. R. Gale |
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| Rok vydání: | 2019 |
| Předmět: |
Cyanobacteria
General Chemical Engineering Genetic Vectors toolkit Computational biology macromolecular substances Polymerase Chain Reaction Fluorescence General Biochemistry Genetics and Molecular Biology transient expression 03 medical and health sciences Synthetic biology Plasmid Escherichia coli Vector (molecular biology) Cloning Molecular Promoter Regions Genetic Synechococcus elongatus UTEX 2973 030304 developmental biology Cloning Synechococcus 0303 health sciences Synechocystis sp. PCC 6803 biology General Immunology and Microbiology 030306 microbiology flow cytometry General Neuroscience Golden Gate Synechocystis plate reader biology.organism_classification Genetically modified organism Transformation (genetics) Conjugation Genetic Expression cassette fluorescence |
| Zdroj: | Gale, G A R, Schiavon Osorio, A A, Puzorjov, A, Wang, B & Mccormick, A J 2019, ' Genetic modification of Cyanobacteria by conjugation using the CyanoGate modular cloning toolkit ', Journal of Visualized Experiments, no. 152, e60451 . https://doi.org/10.3791/60451 |
| ISSN: | 1940-087X |
| DOI: | 10.3791/60451-v |
| Popis: | Cyanobacteria are a diverse group of prokaryotic photosynthetic organisms that can be genetically modified for the renewable production of useful industrial commodities. Recent advances in synthetic biology have led to development of several cloning toolkits such as CyanoGate, a standardized modular cloning system for building plasmid vectors for subsequent transformation or conjugal transfer into cyanobacteria. Here we outline a detailed method for assembling a self-replicating vector (e.g., carrying a fluorescent marker expression cassette) and conjugal transfer of the vector into the cyanobacterial strains Synechocystis sp. PCC 6803 or Synechococcus elongatus UTEX 2973. In addition, we outline how to characterize the performance of a genetic part (e.g., a promoter) using a plate reader or flow cytometry. |
| Databáze: | OpenAIRE |
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