https://scholars.lib.ntu.edu.tw/handle/123456789/390318
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.author | Lin, Y.-J. | en_US |
dc.contributor.author | Liu, W.-T. | en_US |
dc.contributor.author | Stark, H. | en_US |
dc.contributor.author | Huang, C.-T. | en_US |
dc.contributor.author | CHING-TSAN HUANG | en_US |
dc.creator | Lin, Y.-J.;Liu, W.-T.;Stark, H.;Huang, C.-T. | - |
dc.date.accessioned | 2018-09-10T15:18:04Z | - |
dc.date.available | 2018-09-10T15:18:04Z | - |
dc.date.issued | 2015 | - |
dc.identifier.uri | http://www.scopus.com/inward/record.url?eid=2-s2.0-84937966212&partnerID=MN8TOARS | - |
dc.identifier.uri | http://scholars.lib.ntu.edu.tw/handle/123456789/390318 | - |
dc.description.abstract | No commercial vaccines are currently available for enterovirus 71 (EV71) infection. Oral virus-like particle (VLP) vaccines are regarded as a better choice for prevention from food-borne diseases compared with injected whole virus vaccines. Unfortunately, the application of oral VLP vaccines produced from transgenic plants was limited due to the concerns of gene contamination. Alternatively, using transgenic mushrooms retains the advantages of transgenic plants and tremendously reduce risks of gene contamination. Polycistronic expression vectors harboring the glyceraldehyde-3-phospho-dehydrogenase promoter to codrive EV71 structural protein P1 and protease 3C using the 2A peptide of porcine teschovirus-1 were constructed and introduced into Flammulina velutipes via Agrobacterium tumefaciens-mediated transformation. The analyses of the genomic PCR, Southern blotting, and RT-PCR showed that the genes of P1 and 3C were integrated into the chromosomal DNA through a single insertion, and their resulting mRNAs were transcribed. The Western blotting analysis combined with LC-MS/MS demonstrated that EV71 VLPs were composed of the four subunit proteins digested from P1 polyprotein by 3C protease. Through the use of a single particle electron microscope, images of 1705 particles with diameter similar to the EV71 viron were used for 3D reconstruction. Protrusions were observed on the surface in the 2D class averages, and a 3D reconstruction of the VLPs was obtained. In conclusion, EV71 VLPs were successfully produced in transgenic F. velutipes using a polycistronic expression strategy, which indicates that this approach is promising for the development of oral vaccines produced in mushrooms. ? 2015, Springer-Verlag Berlin Heidelberg. | - |
dc.language | en | en |
dc.relation.ispartof | Applied Microbiology and Biotechnology | en_US |
dc.source | AH | - |
dc.subject | 2A peptides; Agrobacterium tumefaciens-mediated transformation; Enterovirus 71; Flammulina velutipes; Polycistron | - |
dc.subject.classification | [SDGs]SDG3 | - |
dc.subject.other | Fungi; Genes; Genetically modified plants; Peptides; Polymerase chain reaction; Proteins; Viruses; 2A Peptide; Agrobacterium tumefaciens-mediated transformation; Enterovirus 71; Flammulina velutipes; Polycistron; Vaccines; oxidoreductase; polyprotein; potassium dihydrogen phosphate; structural protein; virosome; virus protein; biotechnology; chromosome; gene expression; gene flow; genome; mushroom; peptide; polymerase chain reaction; transgenic plant; vaccine; virus; Agrobacterium; Agrobacterium tumefaciens; Article; electron microscopy; Enterovirus 71; enzyme linked immunosorbent assay; Flammulina velutipes; genomics; image processing; immunogenicity; nonhuman; protein processing; reverse transcription polymerase chain reaction; site directed mutagenesis; Southern blotting; virus like agent; Western blotting; Enterovirus A; Flammulina; gene expression profiling; genetic transformation; genetics; liquid chromatography; metabolism; polymerase chain reaction; real time polymerase chain reaction; tandem mass spectrometry; Teschovirus; three dimensional imaging; Agrobacterium tumefaciens; Basidiomycota; Flammulina velutipes; Human enterovirus 71; Porcine teschovirus 1; Agrobacterium tumefaciens; Blotting, Southern; Blotting, Western; Chromatography, Liquid; Enterovirus A, Human; Flammulina; Gene Expression Profiling; Imaging, Three-Dimensional; Microscopy, Electron; Polymerase Chain Reaction; Real-Time Polymerase Chain Reaction; Tandem Mass Spectrometry; Teschovirus; Transformation, Genetic; Viral Proteins; Virosomes | - |
dc.title | Expression of enterovirus 71 virus-like particles in transgenic enoki (Flammulina velutipes) | - |
dc.type | journal article | en |
dc.identifier.doi | 10.1007/s00253-015-6588-z | - |
dc.relation.pages | 6765-6774 | - |
dc.relation.journalvolume | 99 | - |
dc.relation.journalissue | 16 | - |
item.fulltext | no fulltext | - |
item.grantfulltext | none | - |
item.openairetype | journal article | - |
item.openairecristype | http://purl.org/coar/resource_type/c_6501 | - |
item.cerifentitytype | Publications | - |
crisitem.author.dept | Biochemical Science and Technology | - |
crisitem.author.orcid | 0000-0003-1019-784X | - |
crisitem.author.parentorg | College of Life Science | - |
顯示於: | 生化科技學系 |
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