Self-fusion and fusion cell isolation of transformants derived from white rot fungus Phanerochaete sordida YK-624 by simple visual method.
Identifieur interne : 000050 ( Main/Corpus ); précédent : 000049; suivant : 000051Self-fusion and fusion cell isolation of transformants derived from white rot fungus Phanerochaete sordida YK-624 by simple visual method.
Auteurs : Toshio Mori ; Ojiro Kondo ; Tomoki Sumiya ; Hirokazu Kawagishi ; Hirofumi HiraiSource :
- Journal of bioscience and bioengineering [ 1347-4421 ] ; 2020.
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : Fungal Proteins.
- chemical , metabolism : Fungal Proteins, Recombinant Proteins.
- cytology : Phanerochaete.
- genetics : Phanerochaete.
- isolation & purification : Phanerochaete.
- metabolism : Phanerochaete.
- methods : Cell Separation.
Abstract
In order to develop a simple method for crossing two transformants, we first attempted to elucidate the fusion type (self-compatibility or -incompatible) of Phanerochaete sordida YK-624. Two transformants expressing green or red fluorescent protein derived from an auxotrophic mutant were constructed. Each recombinant protein fluoresced by expression as a fused protein with glyceraldehyde-3-phosphate dehydrogenase. On co-culture of both transformants, a number of sequential hyphal cells emitting dual fluorescence were formed at the contact areas of both hyphae. Some of the single cells isolated as protoplasts and chlamydospore from the co-cultures also expressed these fluorescent proteins. These results suggest that P. sordida YK-624 possesses a self-compatible fusion system. In addition, transformant strains with different fluorescence derived from this fungus can readily undergo self-fusion and nuclear interchange events by confrontational and mixed cultivation, and we developed a simple method that allows fused cells to be isolated as chlamydospores.
DOI: 10.1016/j.jbiosc.2019.08.011
PubMed: 31506244
Links to Exploration step
pubmed:31506244Le document en format XML
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<front><div type="abstract" xml:lang="en">In order to develop a simple method for crossing two transformants, we first attempted to elucidate the fusion type (self-compatibility or -incompatible) of Phanerochaete sordida YK-624. Two transformants expressing green or red fluorescent protein derived from an auxotrophic mutant were constructed. Each recombinant protein fluoresced by expression as a fused protein with glyceraldehyde-3-phosphate dehydrogenase. On co-culture of both transformants, a number of sequential hyphal cells emitting dual fluorescence were formed at the contact areas of both hyphae. Some of the single cells isolated as protoplasts and chlamydospore from the co-cultures also expressed these fluorescent proteins. These results suggest that P. sordida YK-624 possesses a self-compatible fusion system. In addition, transformant strains with different fluorescence derived from this fungus can readily undergo self-fusion and nuclear interchange events by confrontational and mixed cultivation, and we developed a simple method that allows fused cells to be isolated as chlamydospores.</div>
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