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(Reference retrieved automatically from SciELO through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Author(s):
Luísa Czamanski Nora [1] ; Relber Aguiar Gonçales [2] ; Leonardo Martins-Santana [3] ; Beatriz Henriques Ferreira [4] ; Fernando Rodrigues [5] ; Rafael Silva-Rocha [6]
Total Authors: 6
Affiliation:
[1] Universidade de São Paulo. Departmento de Biologia Celular e Molecular Biology e Bioagentes Patogênicos. Systems and Synthetic Biology Laboratory - Brasil
[2] Universidade de São Paulo. Departmento de Biologia Celular e Molecular Biology e Bioagentes Patogênicos. Immunochemistry and Glycobiology Laboratory - Brasil
[3] Universidade de São Paulo. Departmento de Biologia Celular e Molecular Biology e Bioagentes Patogênicos. Systems and Synthetic Biology Laboratory - Brasil
[4] University of Minho. Life and Health Sciences Research Institute (ICVS). School of Health Sciences - Portugal
[5] University of Minho. Life and Health Sciences Research Institute (ICVS). School of Health Sciences - Portugal
[6] Universidade de São Paulo. Departmento de Biologia Celular e Molecular Biology e Bioagentes Patogênicos. Systems and Synthetic Biology Laboratory - Brasil
Total Affiliations: 6
Document type: Journal article
Source: GENETICS AND MOLECULAR BIOLOGY; v. 42, n. 2, p. 395-398, 2019-06-13.
Abstract

Abstract 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. (AU)

FAPESP's process: 16/03763-3 - Unveiling the mechanisms of gene regulation at the single cell level in Trichoderma reesei
Grantee:Luísa Czamanski Nora
Support Opportunities: Scholarships in Brazil - Master
FAPESP's process: 16/01946-3 - Construction of synthetic promoters for the expression of cellulases using novel cis-regulatory elements in Trichoderma reesei
Grantee:Leonardo Martins Santana
Support Opportunities: Scholarships in Brazil - Master
FAPESP's process: 12/22921-8 - Synthetic biology approaches for deciphering the logic of signal integration in complex bacterial promoters
Grantee:Rafael Silva Rocha
Support Opportunities: Research Grants - Young Investigators Grants
FAPESP's process: 14/22561-7 - Overexpression of paracoccin: phenotype and virulence of Paracoccidioides brasiliensis strains
Grantee:Relber Aguiar Gonçales
Support Opportunities: Scholarships in Brazil - Doctorate