Brick into the Gateway (BiG): A novel approach for faster cloning combining Golden Gate and Gateway methods.

Autor: Ferigolo LF; Laboratory of Molecular Genetics of Plant Development, Escola Superior de Agricultura 'Luiz de Queiroz' (ESALQ), University of São Paulo, 13418-900 Piracicaba, São Paulo, Brazil., Vicente MH; Laboratory of Molecular Genetics of Plant Development, Escola Superior de Agricultura 'Luiz de Queiroz' (ESALQ), University of São Paulo, 13418-900 Piracicaba, São Paulo, Brazil., Nogueira FTS; Laboratory of Molecular Genetics of Plant Development, Escola Superior de Agricultura 'Luiz de Queiroz' (ESALQ), University of São Paulo, 13418-900 Piracicaba, São Paulo, Brazil. Electronic address: ftsnogue@usp.br.
Jazyk: angličtina
Zdroj: Plasmid [Plasmid] 2022 May; Vol. 121, pp. 102630. Date of Electronic Publication: 2022 Apr 08.
DOI: 10.1016/j.plasmid.2022.102630
Abstrakt: Gateway system is one of the most known cloning systems, which makes it compatible with several expression vectors, including those used for Yeast Two-Hybrid (Y2H) and Bimolecular Fluorescence Complementation (BiFC) assays. However, this system is laborious and expensive due to its two-step cloning. In this research, we developed a new cloning strategy named Brick into the Gateway (BiG). This approach uses GoldenBraid/Gate assemblies to create a DNA fragment of interest flanked by attL sites, which can be directly recombined into Gateway destination vectors. BiG method showed a high recombination efficiency and ensured the correct reading frame, which was successfully tested in Y2H and BiFC assays. BiG has proven to be a rapid, low-cost, reusable, and directional cloning method which allows the merged use of systems.
(Copyright © 2022 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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