Expression of avian reovirus sigmaC protein in transgenic plants.
Avian reovirus (ARV) structural protein, sigmaC encoded by S1 genome segment, is the prime candidate to become a vaccine against ARV infection. Two plant nuclear expression vectors with expression of sigmaC-encoding gene driven by CaMV 35S promoter and rice actin promoter were constructed, respectively. Agrobacterium containing the S1 expression constructs were used to transform alfalfa, and transformants were selected using hygromysin. The integration of S1 transgene in alfalfa chromosome was confirmed by PCR and histochemical GUS staining. Western blot analysis using antiserum against sigmaC was carried out to determine the expression of sigmaC protein in transgenic alfalfa cells. The highest expression levels of sigmaC protein in the cellular extracts of selected p35S-S1 and pAct1-S1 transgenic alfalfa lines were 0.008% and 0.007% of the total soluble protein, respectively. The transgenic alfalfa cells with expression of sigmaC protein pave the way for the development of edible vaccine.
Huang
Liang-Kai
LK
Institute of Biotechnology, National Cheng Kung University, Tainan 701, Taiwan.
Liao
Sin-Chung
SC
Chang
Ching-Chun
CC
Liu
Hung-Jen
HJ
D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods*; D016679:Genome, Viral; D000455:Medicago sativa / Q000235:genetics / Q000378:metabolism*; D030101:Orthoreovirus, Avian / Q000737:chemistry* / Q000235:genetics; D030821:Plants, Genetically Modified / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012231:Rhizobium / Q000235:genetics; D014170:Transformation, Genetic
D036022:Capsid Proteins; D011994:Recombinant Proteins; C031728:sigma 1 protein, reovirus