Characterization of tumor necrosis factor receptor-associated factor gene from megalocytivirus isolated in marine fish
碩士 === 國立臺灣大學 === 漁業科學研究所 === 99 === Viruses belonging to the genus Megalocytivirus in the family Iridoviridae are one of the major agents causing mass mortalities in marine and freshwater fish in Asian countries. Infectious spleen and kidney necrosis virus (ISKNV) is the type species of the Megaloc...
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| Format: | Others |
| Language: | zh-TW |
| Published: |
2011
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| Online Access: | http://ndltd.ncl.edu.tw/handle/78668783583591528375 |
| Summary: | 碩士 === 國立臺灣大學 === 漁業科學研究所 === 99 === Viruses belonging to the genus Megalocytivirus in the family Iridoviridae are one of the major agents causing mass mortalities in marine and freshwater fish in Asian countries. Infectious spleen and kidney necrosis virus (ISKNV) is the type species of the Megalocytivirus genus. ISKNV ORF111L encodes a 296-residue protein that has some similarity to the tumor necrosis factor receptor-associated factor (TRAF) proteins of Homo sapiens and Mus musculus. In this study, a virus isolated from a persistently infected red sea bream fin cell line (PI-RSBF-2) was classified to ISKNV, and we elucidate the viral protein which involved in apoptosis process or cell proliferarion regulation by cloning ISKNV ORF111L and functional assays. Cloned gene sequence revealed 100% identity to ISKNV ORF111L sequence and the deduced amino acid sequences showed high similarity (60-92%) to other Megalocytiviruses TRAF homolog. ISKNV ORF111L contains a TRAF superfamily domain and a RING finger domain but has no transmembrane fragments. The results from phylogenetic analysis shows that ISKNV ORF111L and TRAF homologs from other Megalocytiviruses are in a monophyletic group and this group is relative close to that from fish TRAF homologs. Furthermore, the transcription of ISKNV ORF111L was detected as early as 2 h post infection in ISKNV infected red sea bream fin cells (RSBF-2). Whereas overexpression of ISKNV ORF111L resulted in advanced apoptosis induced by Staurosporine. In cell growth experiment showed that overexpression of ISKNV ORF111L can promote the proliferation RSBF-2 cells.
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