Modular Cloning by Golden Gate Assembly and Possible Application in Pathway Design
Preparation of expression vectors using conventional cloning strategies is laborious and not suitable for the design of metabolic pathways or enzyme cascades, which usually requires the preparation of a vector library to identify productive clones. Recently, Modular Cloning as a novel cloning techn...
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| Format: | Article |
| Language: | deu |
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Swiss Chemical Society
2023-06-01
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| Series: | CHIMIA |
| Subjects: | |
| Online Access: | https://www.chimia.ch/chimia/article/view/6961 |
| _version_ | 1797794113328775168 |
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| author | Zrinka Raguz Nakic Christin Peters |
| author_facet | Zrinka Raguz Nakic Christin Peters |
| author_sort | Zrinka Raguz Nakic |
| collection | DOAJ |
| description |
Preparation of expression vectors using conventional cloning strategies is laborious and not suitable for the design of metabolic pathways or enzyme cascades, which usually requires the preparation of a vector library to identify productive clones. Recently, Modular Cloning as a novel cloning technique in synthetic biology has been developed. Modular Cloning relies on Golden Gate assembly and supports preparation of individual expression vectors in one-step and one-pot reactions, thus allowing rapid generation of vector libraries. A number of Modular Cloning toolkits for specific applications has been established, providing a collection of distinct genetic elements such as promoters, ribosome binding sites and tags, that can be combined individually in one-step using defined fusion sites. Modular Cloning has been successfully applied to generate various strains for producing value-added compounds. This was achieved by orchestrating complex pathways involving up to 20 enzymes. Due to the novelty of the genetic approach, industrial applications are still rare. In addition, some applications are limited due to the lack of high-throughput screening methods. This shifts the bottleneck from library preparation to screening capacity and needs to be addressed by future developments to pave the path for the establishment of Modular Cloning in industrial applications.
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| first_indexed | 2024-03-13T02:58:04Z |
| format | Article |
| id | doaj.art-e18c6a370b7b476ebdb28fa02924a28c |
| institution | Directory Open Access Journal |
| issn | 0009-4293 2673-2424 |
| language | deu |
| last_indexed | 2024-03-13T02:58:04Z |
| publishDate | 2023-06-01 |
| publisher | Swiss Chemical Society |
| record_format | Article |
| series | CHIMIA |
| spelling | doaj.art-e18c6a370b7b476ebdb28fa02924a28c2023-06-28T03:42:17ZdeuSwiss Chemical SocietyCHIMIA0009-42932673-24242023-06-0177610.2533/chimia.2023.437Modular Cloning by Golden Gate Assembly and Possible Application in Pathway Design Zrinka Raguz Nakic0Christin Peters1Biosystems Technology Group, Institute of Chemistry and Biotechnology, Zurich University of Applied Sciences, CH-8820 WädenswilBiosystems Technology Group, Institute of Chemistry and Biotechnology, Zurich University of Applied Sciences, CH-8820 Wädenswil Preparation of expression vectors using conventional cloning strategies is laborious and not suitable for the design of metabolic pathways or enzyme cascades, which usually requires the preparation of a vector library to identify productive clones. Recently, Modular Cloning as a novel cloning technique in synthetic biology has been developed. Modular Cloning relies on Golden Gate assembly and supports preparation of individual expression vectors in one-step and one-pot reactions, thus allowing rapid generation of vector libraries. A number of Modular Cloning toolkits for specific applications has been established, providing a collection of distinct genetic elements such as promoters, ribosome binding sites and tags, that can be combined individually in one-step using defined fusion sites. Modular Cloning has been successfully applied to generate various strains for producing value-added compounds. This was achieved by orchestrating complex pathways involving up to 20 enzymes. Due to the novelty of the genetic approach, industrial applications are still rare. In addition, some applications are limited due to the lack of high-throughput screening methods. This shifts the bottleneck from library preparation to screening capacity and needs to be addressed by future developments to pave the path for the establishment of Modular Cloning in industrial applications. https://www.chimia.ch/chimia/article/view/6961Expression controlGolden Gate assemblyModular CloningPathway design |
| spellingShingle | Zrinka Raguz Nakic Christin Peters Modular Cloning by Golden Gate Assembly and Possible Application in Pathway Design CHIMIA Expression control Golden Gate assembly Modular Cloning Pathway design |
| title | Modular Cloning by Golden Gate Assembly and Possible Application in Pathway Design |
| title_full | Modular Cloning by Golden Gate Assembly and Possible Application in Pathway Design |
| title_fullStr | Modular Cloning by Golden Gate Assembly and Possible Application in Pathway Design |
| title_full_unstemmed | Modular Cloning by Golden Gate Assembly and Possible Application in Pathway Design |
| title_short | Modular Cloning by Golden Gate Assembly and Possible Application in Pathway Design |
| title_sort | modular cloning by golden gate assembly and possible application in pathway design |
| topic | Expression control Golden Gate assembly Modular Cloning Pathway design |
| url | https://www.chimia.ch/chimia/article/view/6961 |
| work_keys_str_mv | AT zrinkaraguznakic modularcloningbygoldengateassemblyandpossibleapplicationinpathwaydesign AT christinpeters modularcloningbygoldengateassemblyandpossibleapplicationinpathwaydesign |