| Summary: | 碩士 === 國立臺灣大學 === 漁業科學研究所 === 106 === Nervous necrosis virus ( NNV) is a positive sense single-stranded RNA virus, it infects more than 50 species of marine fish and causes significant economic losses. To investigate the interaction of NNV coat protein and grouper brain ( GB) cell, we set up the NNV virus-like particle-coupled affinity column. By LC-MS/MS analysis, we define the 30 kDa band in SDS-polyacrylamide gel is ribosomal protein L7a (RPL7a). The interaction between NNV coat protein and RPL7a were further confirmed by virus-overlay protein binding assay ( VOPBA), and the binding region of RPL7a against NNV coat protein was defined to 1-85 a.a. By cotransfection of expression constructs which encoded NNV coat protein and RPL7a with HA or Myc tags, we find that both NNV coat protein and RPL7a colocalized in nucleolus 15 hr after cotransfection . With NNV infection, recombinant RPL7a was found to translocate from nucleolus to cytoplasm. The protein level of RPL7a in GB cell was constant during NNV infection by Western analysis. However, the amount of RNA2 in NNV infected GB cells was increase slightly after RPL7a knockdown by RNAi. Taken together, these results showed that the interaction between NNV coat protein and RPL7a relocated RPL7a from nucleolus to cytoplasm and influenced the propogation of NNV.
|
|---|
