[Immunogenicity of co-expressed p30-gp90 against reticuloendotheliosis virus].
Ruiai Chen, Yanpeng Li, Kai Guo, Zhengwei Liu
Abstract
Ruiai Chen, Yanpeng Li, Kai Guo, Zhengwei Liu
Abstract
OBJECTIVE: To study the immunogenicity of co-expression of p30, one of a group specific antigens, and glycoprotein gp90 genes of Reticuloendotheliosis virus (REV). METHODS: p30 and gp90 genes were amplified with the template of plasmid pPB101 containing the whole sequence of spleen necrosis virus (SNV), and cloned into pET-28a(+) vector. The positive clone pET-p30-gp90 was identified by enzyme analysis and sequencing. The co-expressed protein p30-gp90 was confirmed by SDS-PAGE and Western blot analysis. After quantitation, the purified protein p30-gp90 was immunized to Balb/c mice three times to prepare the p30-gp90 specific anti-serum. Afterwards, we detected the REV infected Chick Embryo Fibroblasts (CEF) by Immunofluorescence assay (IFA) with the prepared anti-serum. RESULTS: The p30-gp90 was co-expressed efficiently by SDS-PAGE and Western blot analysis. The anti-serum of p30-gp90 reacts with REV infected CEF in IFA. CONCLUSION: The expressed protein of p30-gp90 keeps good immunogenicity.
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OBJECTIVE: To study the immunogenicity of co-expression of p30, one of a group specific antigens, and glycoprotein gp90 genes of Reticuloendotheliosis virus (REV). METHODS: p30 and gp90 genes were amplified with the template of plasmid pPB101 containing the whole sequence of spleen necrosis virus (SNV), and cloned into pET-28a(+) vector. The positive clone pET-p30-gp90 was identified by enzyme analysis and sequencing. The co-expressed protein p30-gp90 was confirmed by SDS-PAGE and Western blot analysis. After quantitation, the purified protein p30-gp90 was immunized to Balb/c mice three times to prepare the p30-gp90 specific anti-serum. Afterwards, we detected the REV infected Chick Embryo Fibroblasts (CEF) by Immunofluorescence assay (IFA) with the prepared anti-serum. RESULTS: The p30-gp90 was co-expressed efficiently by SDS-PAGE and Western blot analysis. The anti-serum of p30-gp90 reacts with REV infected CEF in IFA. CONCLUSION: The expressed protein of p30-gp90 keeps good immunogenicity.
Key concepts: Immunogenicity, Reticuloendotheliosis virus, Biology, Molecular biology, Virology, Virus, Western blot, Antigen