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

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Autor(es):
Ferigolo, Leticia F. ; Vicente, Mateus H. ; Nogueira, Fabio T. S.
Número total de Autores: 3
Tipo de documento: Artigo Científico
Fonte: PLASMID; v. 121, p. 8-pg., 2022-04-12.
Resumo

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. (AU)

Processo FAPESP: 19/20157-8 - Avaliação de possíveis interações entre as vias de florescimento fotoperiódica, age e giberelina com o módulo SP/SFT e seus impactos sobre a arquitetura do tomateiro (Solanum lycopersicum L.)
Beneficiário:Mateus Henrique Vicente
Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
Processo FAPESP: 18/17441-3 - Controle molecular da arquitetura vegetal: interconexão entre microRNAs, fatores de transcrição e fitohormônios
Beneficiário:Fabio Tebaldi Silveira Nogueira
Modalidade de apoio: Auxílio à Pesquisa - Temático