English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Expression of enterovirus 71 virus-like particles in transgenic enoki (Flammulina velutipes).

Lin, Y. J., Liu, W., Stark, H., & Huang, C. T. (2015). Expression of enterovirus 71 virus-like particles in transgenic enoki (Flammulina velutipes). Applied Microbiology and Biotechnology, 99(16), 6765-6774. doi:10.1007/s00253-015-6588-z.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-31DA-C Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-31DD-6
Genre: Journal Article

Files

show Files
hide Files
:
2180533.pdf (Publisher version), 3MB
 
File Permalink:
-
Name:
2180533.pdf
Description:
-
Visibility:
Restricted (Max Planck Institute for Biophysical Chemistry (Karl Friedrich Bonhoeffer Institute), Göttingen; )
MIME-Type / Checksum:
application/pdf
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-

Locators

show

Creators

show
hide
 Creators:
Lin , Y. J., Author
Liu, W.1, Author              
Stark, H.1, Author              
Huang, C. T., Author
Affiliations:
1Research Group of 3D Electron Cryo-Microscopy, MPI for biophysical chemistry, Max Planck Society, ou_578577              

Content

show
hide
Free keywords: Flammulina velutipes; Agrobacterium tumefaciens-mediated transformation; 2A peptides; Polycistron; Enterovirus 71
 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.

Details

show
hide
Language(s): eng - English
 Dates: 2015-05-092015-08
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1007/s00253-015-6588-z
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Applied Microbiology and Biotechnology
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 99 (16) Sequence Number: - Start / End Page: 6765 - 6774 Identifier: -