Overexpression and repression of the tyrosinase gene in Lentinula edodes using the pChG vector
| Autor: | Hisayuki Watanabe, Toshitsugu Sato, Machiko Takahashi, Junji Hasegawa |
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| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
0301 basic medicine Tyrosinase Genetic Vectors Shiitake Mushrooms Bioengineering Biology 01 natural sciences Applied Microbiology and Biotechnology Gene Expression Regulation Enzymologic 03 medical and health sciences Transformation Genetic Gene Expression Regulation Fungal 010608 biotechnology Gene expression Fruiting Bodies Fungal Gene Silencing Promoter Regions Genetic Gene Mycelium Chitin Synthase Organisms Genetically Modified Strain (chemistry) Monophenol Monooxygenase fungi Protoplast biology.organism_classification Molecular biology Up-Regulation Transformation (genetics) 030104 developmental biology Lentinula Biotechnology |
| Zdroj: | Journal of Bioscience and Bioengineering. 128:1-7 |
| ISSN: | 1389-1723 |
| Popis: | Tyrosinase is an industrially useful enzyme, however, it causes gill browning of Lentinula edodes fruiting bodies during preservation. In this study, we constructed two vectors, pChG-gTs and pChG-gTa, expressing sense and antisense tyrosinase gene of L. edodes, respectively, using promoters derived from the glyceraldehyde-3-phosphate dehydrogenase gene. The host strain SR-1 of L. edodes was selected because of its fast growth, high protoplast yield, and high regeneration rate. Upon transformation of the host strain SR-1 with the pChG-gTs vector, a clone with 3.6-fold and 14.5-fold higher tyrosinase activity in vegetative mycelia and in fresh gills, respectively, than that of the host strain was obtained from nine transformants. Similarly, two clones containing the pChG-gTa vector with effectively repressed tyrosinase gene expression in vegetative mycelia and gills during the late stage of post-harvest preservation of fruiting bodies were obtained from 10 transformants. However, it remained unclear whether repression of the tyrosinase gene prevented gill browning, as the host strain also showed less browning than a commercial strain. Thus, this study highlights the usefulness of the pChG vector in expressing homologous enzyme coding genes in the vegetative mycelia and fruiting bodies of L. edodes. |
| Databáze: | OpenAIRE |
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