Synthetic and minimalist vectors for Agrobacterium tumefaciens-mediated transformation of fungi

• Autor(a) principal: Nora, Luísa Czamanski
• Data de Publicação: 2019
• Outros Autores: Gonçales, Relber Aguiar, Martins-Santana, Leonardo, Ferreira, Beatriz Henriques, Rodrigues, Fernando José dos Santos, Silva-Rocha, Rafael
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/1822/62304
• topic: Synthetic biology; Fungi; Agrobacterium tumefaciens; Transformation; Vectors; Science & Technology

We present a collection of minimalist binary vectors for transformation through ATMT applicable to several fungi species. pLUO plasmid binary vectors consist of a reporter module containing fluorescent proteins, mCherry or eGFP, flanked by a multiple cloning site and a transcription terminator site. They also present a synthetic gene allowing resistance to Hygromicin B flanked by alternate promoters, one for yeast and another for filamentous fungi. Left and right borders were added for Agrobacterium tumefaciens recognition, and a minimal broad-host range RK2 replication origin. Transformation was validated in the pathogenic fungus Paracoccidioides lutzii. Hence, we developed an efficient and reliable molecular tool for fungal transformation: minimalist, synthetic, modular, and available in four different versions, and these can still be readily modified using a few primers and few cloning steps.