Expression of avian reovirus minor capsid protein in plants

Shih Wei Lu, Kuo Chih Wang, Hung Jen Liu, Ching Dong Chang, Hao Jen Huang, Ching Chun Chang

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The minor coat protein of the avian reovirus (ARV), σC, encoded by the S1 genome segment, is one of the major candidates for the development of a subunit vaccine against ARV infection. To develop a plant-based vaccine to immunize poultry against ARV infection, we constructed 4 plant nuclear expression vectors with or without codon modification of the S1 gene, and their expression was driven by a CaMV 35S promoter or rice actin1 promoter. In addition, the expressed σC proteins were targeted subcellularly to cytosol or chloroplasts, respectively. Agrobacterium containing the S1 expression constructs was used to transform tobacco leaf disks, and transformants were selected with kanamycin (100 μg/ml). The integration of the S1 transgene into the tobacco chromosome was confirmed by PCR and Southern blot analysis. Western blot analysis with antiserum against σC was performed to determine the expression of σC protein in transgenic tobacco plants. The highest expression levels of σC protein in the cellular extracts of selected p35S1, pActS1 and p35UmS1 transgenic lines were 0.013%, 0.021% and 0.0013% of the total soluble protein, respectively, but the protein was barely detectable in p35TmS1 transgenic lines. However, the level of σC protein expression was not associated with the level of corresponding RNA transcripts in selected transgenic lines. Taken together, the results suggest that the major limiting factor for the expression of σC protein in plants might be at the post-transcriptional level.

Original languageEnglish
Pages (from-to)287-293
Number of pages7
JournalJournal of Virological Methods
Volume173
Issue number2
DOIs
Publication statusPublished - 2011 May 1

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Avian orthoreovirus
Capsid Proteins
Protein C
Reoviridae Infections
Tobacco
Plant Proteins
Agrobacterium
Subunit Vaccines
Kanamycin
Genetically Modified Plants
Chloroplasts
Poultry
Southern Blotting
Transgenes
Codon
Cytosol
Immune Sera
Proteins
Vaccines
Chromosomes

All Science Journal Classification (ASJC) codes

  • Virology

Cite this

Lu, Shih Wei ; Wang, Kuo Chih ; Liu, Hung Jen ; Chang, Ching Dong ; Huang, Hao Jen ; Chang, Ching Chun. / Expression of avian reovirus minor capsid protein in plants. In: Journal of Virological Methods. 2011 ; Vol. 173, No. 2. pp. 287-293.
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Expression of avian reovirus minor capsid protein in plants. / Lu, Shih Wei; Wang, Kuo Chih; Liu, Hung Jen; Chang, Ching Dong; Huang, Hao Jen; Chang, Ching Chun.

In: Journal of Virological Methods, Vol. 173, No. 2, 01.05.2011, p. 287-293.

Research output: Contribution to journalArticle

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