Identification and characterization of Toll-interleukin 1 receptor domain-containing adapter protein（TIRAP）in Atlantic salmon（Salmo salar）

• Main Authors: Lin, Ching-Lun, 林敬倫
• Other Authors: Lee, Kuo-Kau
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/8755ny

碩士 === 國立臺灣海洋大學 === 水產養殖學系 === 106 === Toll-interleukin 1 receptor domain-containing adapter protein(TIRAP)is an important adaptor protein in the pathways of inflammatory response and cytokine secretion, and therefore plays an important role in the innate immunity of fish. In this thesis, full-length cDNA of Atlantic salmon TIRAP gene(SsTIRAP)has been cloned into vectors and sequenced. The sequenced SsTIRAP is identical to the in silico predicted salmon TIRAP provided by the National Center for Biotechnology Information(NCBI). The evolutionary status of SsTIRAP and its relationship to homologous genes from selected species was analyzed. A conserved gene synteny was found with the salmon TIRAP as seen in other vertebrate loci. Furthermore, real-time polymerase chain reaction(RT-PCR)was performed to determine the expression level of SsTIRAP in the liver, spleen, trunk kidney, head kidney, heart, muscle, intestine, skin, brain and gills of Atlantic salmon. For gene expression analysis, the SsTIRAP is expressed at a relatively high level in the spleen, followed by trunk kidney skin, head kidney, muscle, gills, brain, heart and intestine, with the lowest expression level in the liver. Finally, Western blotting and immunofluorescence staining analysis was performed to reveal the intracellular expression of V5 antigen tagged SsTIRAP in the transfected and overexpressed Human embryonic kidney(HEK)293T cells.