A novel improved method for Aspergillus nidulans transformation

Koukaki, Marina; Giannoutsou, Eleni; Karagouni, Amalia D.; Diallinas, George

doi:10.1016/s0167-7012(03)00208-2

Cited by 81 publications

(53 citation statements)

References 17 publications

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“…The pabaA1, riboB2, pyroA4, pyrG89, and argB2 are genetic mutations resulting in auxotrophies for p-aminobenzoic acid, riboflavine, pyridoxin, uracil/uridine and arginine respectively (http:// www.fgsc.net/Aspergillus/gene_list/supplement.html). A. nidulans transformation was performed as described previously (Koukaki et al, 2003). DNA from A. nidulans was extracted as described in http://www.fgsc.net.…”

Section: Methodsmentioning

confidence: 99%

Minos as a novel Tc1/mariner-type transposable element for functional genomic analysis in Aspergillus nidulans

Evangelinos¹,

Anagnostopoulos²,

Karvela-Kalogeraki³

et al. 2015

Fungal Genetics and Biology

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Section: Methodsmentioning

confidence: 99%

Minos as a novel Tc1/mariner-type transposable element for functional genomic analysis in Aspergillus nidulans

Evangelinos¹,

Anagnostopoulos²,

Karvela-Kalogeraki³

et al. 2015

Fungal Genetics and Biology

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“…Media and chemical reagents were obtained from Sigma or AppliChem (Bioline Scientific SA, Hellas, Greece). Transformations were performed as described previously (48). A single-copy plasmid integration of the FcyB served as a standard wild-type control.…”

Section: Methodsmentioning

confidence: 99%

Modeling, Substrate Docking, and Mutational Analysis Identify Residues Essential for the Function and Specificity of a Eukaryotic Purine-Cytosine NCS1 Transporter

Krypotou

Kosti

Amillis

et al. 2012

Journal of Biological Chemistry

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Background:The purine-cytosine FcyB transporter is a prototype member of the NCS1 family. Results: Using homology modeling, substrate docking, and rational mutational analysis, we identify residues critical for function and specificity. Conclusion: Important aspects concerning the molecular mechanism and evolution of transporter specificity are revealed. Significance: The first systematic approach on structure-function-specificity relationships in a eukaryotic NCS1 member is shown.

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“…Hyphal cells and/or germlings that are used to prepare protoplasts of A. nidulans contain multiple nuclei (Fiddy and Trinci 1976), therefore problems associated with the number of nuclei in a cell after protoplast fusion would not be a significant issue for this organism. Therefore, together with controversy regarding the occurrence of endocytosis in filamentous fungi (de Vries et al 2003), fusion could have been considered as a mechanism for extracellular DNA uptake in filamentous fungi (Koukaki et al 2003). However, in S.…”

Section: Introductionmentioning

confidence: 99%

Comparison between polyethylene glycol- and polyethylenimine-mediated transformation of Aspergillus nidulans

2008

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Genetic transformation of many filamentous fungi is carried out by a protocol that utilizes polyethylene glycol (PEG) and calcium ion (Ca 2+ ). This method has remained practically unchanged for more than 20 years, but the roles these molecules play are not definitively understood. To gain a better understanding, we have compared PEG transformation to a protocol using polyethylenimine (PEI) that is the basis for non-viral transfection in mammals and which has a well established molecular model for assisting DNA uptake. Protoplasts of Aspergillus nidulans could be transformed in the presence of Ca 2+ with a relatively high ratio of PEI to DNA molecules. By comparing PEI and PEG in terms of interaction with DNA, fungal protoplasts, and response to different transformation conditions, we propose that the role of PEG is most likely to function after transforming DNA is incorporated into protoplasts, rather than the accepted view that it functions outside of the cell. Confirmation that protoplast fusion was not involved in DNA uptake is consistent with this hypothesis.

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A novel improved method for Aspergillus nidulans transformation

Cited by 81 publications

References 17 publications

Minos as a novel Tc1/mariner-type transposable element for functional genomic analysis in Aspergillus nidulans

Minos as a novel Tc1/mariner-type transposable element for functional genomic analysis in Aspergillus nidulans

Modeling, Substrate Docking, and Mutational Analysis Identify Residues Essential for the Function and Specificity of a Eukaryotic Purine-Cytosine NCS1 Transporter

Comparison between polyethylene glycol- and polyethylenimine-mediated transformation of Aspergillus nidulans

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