Easy method for six-fragment Golden Gate Assembly of modular vectors
Efficient cloning techniques are a requirement for synthetic biology. This study provides a simplified cloning method based on Golden Gate Assembly that can be used for rapid vector construction. The building of multiple expression vectors with customizable modules is achieved in a timely manner wit...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2023-02-01
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| Series: | BioTechniques |
| Subjects: | |
| Online Access: | https://www.future-science.com/doi/10.2144/btn-2022-0041 |
| _version_ | 1826992594818695168 |
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| author | Ionut Adrian Cepleanu-Pascu Miruna Stan Sebastian Cocioba Ileana Stoica |
| author_facet | Ionut Adrian Cepleanu-Pascu Miruna Stan Sebastian Cocioba Ileana Stoica |
| author_sort | Ionut Adrian Cepleanu-Pascu |
| collection | DOAJ |
| description | Efficient cloning techniques are a requirement for synthetic biology. This study provides a simplified cloning method based on Golden Gate Assembly that can be used for rapid vector construction. The building of multiple expression vectors with customizable modules is achieved in a timely manner with minimal hands-on time by removing unnecessary steps in the workflow. The authors constructed a total of 21 mammalian episomal expression vectors and conducted a fluorescence expression comparison for different regulatory region combinations post-transfection in HEK293T and HEPG2 cells. Screening revealed that using the EF-1α promoter in combination with the bovine growth hormone polyadenylation sequence seemed to perform best in the types of cells tested compared with other variants. |
| first_indexed | 2024-04-10T06:29:41Z |
| format | Article |
| id | doaj.art-937a6ea49ddf43c79633285ee4b65d3a |
| institution | Directory Open Access Journal |
| issn | 0736-6205 1940-9818 |
| language | English |
| last_indexed | 2025-02-18T08:52:59Z |
| publishDate | 2023-02-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj.art-937a6ea49ddf43c79633285ee4b65d3a2024-11-02T23:57:16ZengTaylor & Francis GroupBioTechniques0736-62051940-98182023-02-01742859910.2144/btn-2022-0041Easy method for six-fragment Golden Gate Assembly of modular vectorsIonut Adrian Cepleanu-Pascu0Miruna Stan1Sebastian Cocioba2Ileana Stoica31Department of Genetics, Faculty of Biology, University of Bucharest, Bucharest, Romania2Department of Biochemistry & Molecular Biology, Faculty of Biology, University of Bucharest, Bucharest, Romania3Binomica Labs, Long Island City, New York, NY 11101, USA1Department of Genetics, Faculty of Biology, University of Bucharest, Bucharest, RomaniaEfficient cloning techniques are a requirement for synthetic biology. This study provides a simplified cloning method based on Golden Gate Assembly that can be used for rapid vector construction. The building of multiple expression vectors with customizable modules is achieved in a timely manner with minimal hands-on time by removing unnecessary steps in the workflow. The authors constructed a total of 21 mammalian episomal expression vectors and conducted a fluorescence expression comparison for different regulatory region combinations post-transfection in HEK293T and HEPG2 cells. Screening revealed that using the EF-1α promoter in combination with the bovine growth hormone polyadenylation sequence seemed to perform best in the types of cells tested compared with other variants.https://www.future-science.com/doi/10.2144/btn-2022-0041cloningGolden GateGolden Gate standardmammalian expression vectorsS/MARs |
| spellingShingle | Ionut Adrian Cepleanu-Pascu Miruna Stan Sebastian Cocioba Ileana Stoica Easy method for six-fragment Golden Gate Assembly of modular vectors BioTechniques cloning Golden Gate Golden Gate standard mammalian expression vectors S/MARs |
| title | Easy method for six-fragment Golden Gate Assembly of modular vectors |
| title_full | Easy method for six-fragment Golden Gate Assembly of modular vectors |
| title_fullStr | Easy method for six-fragment Golden Gate Assembly of modular vectors |
| title_full_unstemmed | Easy method for six-fragment Golden Gate Assembly of modular vectors |
| title_short | Easy method for six-fragment Golden Gate Assembly of modular vectors |
| title_sort | easy method for six fragment golden gate assembly of modular vectors |
| topic | cloning Golden Gate Golden Gate standard mammalian expression vectors S/MARs |
| url | https://www.future-science.com/doi/10.2144/btn-2022-0041 |
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