A Fruiting Body Tissue Method for Efficient
            <i>Agrobacterium</i>
            -Mediated Transformation of
            <i>Agaricus bisporus</i>

Chen, Xi; Stone, Michelle C.; Schlagnhaufer, Carl D.; Romaine, C. P.

doi:10.1128/aem.66.10.4510-4513.2000

Cited by 217 publications

(163 citation statements)

References 14 publications

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“…Until now, there was no available method for the efficient transformation of human gene into mushroom using Agrobacterium; pioneering efforts were made by De Groot et al [15] and Chen et al [16] for Agrobacterium-mediated transformation of fungi. In this study, we explored a different approach to transform human gene into P. eryngii mediated by Agrobacterium.…”

Section: Discussionmentioning

confidence: 99%

“…Further developments in Agrobacterium tumefaciens-mediated transformation of fungi have been reported [16] and several studies continued to optimize the technology. In the present study, we introduced for the first time the hGH2 gene into the edible mushroom Pleurotus eryngii, using Agrobacterium-mediated transformation, which resulted in a successful expression of the human gene.…”

Section: Introductionmentioning

confidence: 99%

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Expression of human growth hormone gene in Pleurotus eryngii

Kim

Sapkota

Choi³

et al. 2010

Open Life Sciences

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Recombinant human growth hormone (rhGH) has been widely used in many medical applications. Large scale production of rhGH is important for a wider application of this molecule in the field of proteomics. This investigation reports a modified Agrobacterium-mediated method for transfer and expression of human growth hormone gene in the popular edible mushroom Pleurotus eryngii. A binary vector pCAMBIA1304 containing the hGH2 gene was constructed and introduced into Pleurotus eryngii via Agrobacterium tumefaciens-mediated transformation. Integration of hGH2 into mushroom genome was detected by PCR and expression was confirmed by Western blot assay. The successful expression of the human growth hormone gene in mushroom suggests that the proposed modified transformation system is probably useful for the production of transgenic mushrooms.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Expression of human growth hormone gene in Pleurotus eryngii

Kim

Sapkota

Choi³

et al. 2010

Open Life Sciences

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“…Nuclear accumulation of free GFP is typical for all eukaryotic organisms as GFP is a small protein, which is below the exclusion size for passive transport through the nuclear pore complex, in contrast to proteasomes (Fabre and Hurt 1994). In previous studies, Chen et al (2000) could efficiently transform Agaricus bisporus with the pBGgHg vector that was used in the present study, but were unable to detect any GFP fluorescence. Moreover, using the same vector, Combier et al (2003) were able to obtain the integration of the EGFP construct as part of the T-DNA into the H. cylindrosporum genome but did not observe GFP fluorescence.…”

Section: Microscopic Fluorescence Analysismentioning

confidence: 51%

“…The binary plasmid pBGgHg (Chen et al 2000) was kindly provided by C. Peter Romaine (The Pennsylvania State University). This vector consists of a pCAMBIA1300 backbone containing the Escherichia coli hygromycin B phosphotransferase (hph) and the EGFP genes, both under the control of the Agaricus bisporus glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter and the cauliflower mosaic virus (CaMV) 35S terminator.…”

Section: Strains Media and Plasmidsmentioning

confidence: 99%

Functional expression of the green fluorescent protein in the ectomycorrhizal model fungus Hebeloma cylindrosporum

et al. 2006

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Hebeloma cylindrosporum is a model fungus for mycorrhizal studies because of its fast growth rate, simple nutritional requirements, and completion of its life cycle in vitro, and because it is amenable to transformation. To advance cell biological research during establishment of symbiosis, a tool that would enable the direct visualisation of fusion proteins in the different symbiotic tissues [namely, the expression of reporter genes such as Green Fluorescent Protein (GFP)] was still a missing tool. In the present study, H. cylindrosporum was transformed using Agrobacterium carrying the binary plasmid pBGgHg containing the Escherichia coli hygromycin B phosphotransferase (hph) and the EGFP genes, both under the control of the Agaricus bisporus glyceraldehyde-3-phosphate dehydrogenase promoter. EGFP expression was successfully detected in transformants. The fluorescence was uniformly distributed in the hyphae, while no significant background signal was detected in control hyphae. The suitability of EGFP for reporter gene studies in Hebeloma cylindrosporum was demonstrated opening up new perspectives in the Hebeloma genetics.

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“…In order to investigate whether the promoter type could influence the transformation efficiency in M. perniciosa, the gpd promoters from A. bisporus (pBHg and pHCP) and A. nidulans (pAN7-1) were tested. The plasmid pBHg was used initially for Agrobacterium-mediated fungal transformation (Chen et al, 2000).The plasmid pHCP constructed in this work originated from pBHg. The transformation efficiency with circular pAN7-1 was 4 transformants/µg and the first transformant appeared after two weeks.…”

Section: Resultsmentioning

confidence: 99%