Expression of the nucleocapsid protein of Porcine Reproductive and Respiratory Syndrome Virus in soybean seed yields an immunogenic antigenic protein
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Porcine reproductive and respiratory syndrome (PRRS), caused by the PRRS virus (PRRSV), is a serious disease of swine and contributes to severe worldwide economic losses in swine production. Current vaccines against PRRS rely on the use of an attenuated-live virus; however, these are unreliable. Thus, alternative effective vaccines against PRRS are needed. Plant-based subunit vaccines offer viable, safe, and environmentally friendly alternatives to conventional vaccines. In this study, efforts have been undertaken to develop a soybean-based vaccine against PRRSV. A construct carrying a synthesized PRRSV-ORF7 antigen, nucleocapsid N protein of PRRSV, has been introduced into soybean, Glycine max (L.) Merrill. cvs. Jack and Kunitz, using Agrobacterium-mediated transformation. Transgenic plants carrying the sORF7 transgene have been successfully generated. Molecular analyses of T 0 plants confirmed integration of the transgene and transcription of the PRRSV-ORF7. Presence of a 15-kDa protein in seeds of T 1 transgenic lines was confirmed by Western blot analysis using PRRSV-ORF7 antisera. The amount of the antigenic protein accumulating in seeds of these transgenic lines was up to 0. 65%25 of the total soluble protein (TSP). A significant induction of a specific immune response, both humoral and mucosal, against PRRSV-ORF7 was observed following intragastric immunization of BALB/c female mice with transgenic soybean seeds. These findings provide a %27proof of concept%27, and serve as a critical step in the development of a subunit plant-based vaccine against PRRS. © 2011 Springer-Verlag.
Porcine reproductive and respiratory syndrome (PRRS), caused by the PRRS virus (PRRSV), is a serious disease of swine and contributes to severe worldwide economic losses in swine production. Current vaccines against PRRS rely on the use of an attenuated-live virus; however, these are unreliable. Thus, alternative effective vaccines against PRRS are needed. Plant-based subunit vaccines offer viable, safe, and environmentally friendly alternatives to conventional vaccines. In this study, efforts have been undertaken to develop a soybean-based vaccine against PRRSV. A construct carrying a synthesized PRRSV-ORF7 antigen, nucleocapsid N protein of PRRSV, has been introduced into soybean, Glycine max (L.) Merrill. cvs. Jack and Kunitz, using Agrobacterium-mediated transformation. Transgenic plants carrying the sORF7 transgene have been successfully generated. Molecular analyses of T 0 plants confirmed integration of the transgene and transcription of the PRRSV-ORF7. Presence of a 15-kDa protein in seeds of T 1 transgenic lines was confirmed by Western blot analysis using PRRSV-ORF7 antisera. The amount of the antigenic protein accumulating in seeds of these transgenic lines was up to 0. 65%25 of the total soluble protein (TSP). A significant induction of a specific immune response, both humoral and mucosal, against PRRSV-ORF7 was observed following intragastric immunization of BALB/c female mice with transgenic soybean seeds. These findings provide a 'proof of concept', and serve as a critical step in the development of a subunit plant-based vaccine against PRRS. © 2011 Springer-Verlag.
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Immunogenicity; Plant-based vaccine; PRRS viral subunit; Transgenic soybean nucleocapsid protein; virus antigen; animal; Arterivirus; article; Bagg albino mouse; enzyme linked immunosorbent assay; female; genetics; immunology; metabolism; mouse; plant seed; reverse transcription polymerase chain reaction; soybean; transgenic plant; Animals; Antigens, Viral; Enzyme-Linked Immunosorbent Assay; Female; Mice; Mice, Inbred BALB C; Nucleocapsid Proteins; Plants, Genetically Modified; Porcine respiratory and reproductive syndrome virus; Reverse Transcriptase Polymerase Chain Reaction; Seeds; Soybeans; Agrobacterium; Arterivirus; Glycine max; Mus; Porcine respiratory and reproductive syndrome virus; Rhizobium; Suidae; Sus
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