Expression analysis of white spot syndrome virus (WSSV) late gene p150

• Main Authors: Yi-Hsing Lin, 林薏欣
• Other Authors: Guang-Hsiung Kou
• Format: Others
• Language: zh-TW
• Published: 2002
• Online Access: http://ndltd.ncl.edu.tw/handle/87162558602676987185

碩士 === 國立臺灣大學 === 動物學研究所 === 90 === One of the white spot syndrome virus (WSSV) genes, p150, that encodes an acidic protein was identified and sequenced from the predicted open reading frame, WSSV067, from the Taiwan isolate. The p150 gene that contains 3,906 nucleotides potentially encodes a protein of 1,301 amino acids with two glutamic acid-rich domains in the N-terminus and three transmembrane regions in the C-terminus. The predicted molecular weight of P150 is 143.9 kDa. The transcript of p150, which was detected at 12 hr post-infection and reached a maximal level at 18 hr, belonged to a late gene of WSSV. The mRNA size of p150 gene is about 4.3 kb, and the poly A addition site is 15 bp downstream of the poly A signal. When compare the region between poly A signal and poly A addition site of known WSSV genes of p150, rr1, DNApol and the predicted open reading frame WSSV202 and WSSV217, a consensus sequence was found, (T/A)ACATATA(T/A)A(T/A). The product of in vitro transcription and translation of p150 gene was detected as a 150 kDa protein. Immunofluorescent staining of P150 protein in insect cell line showed that the P150 localized in cytoplasm, and might bind to some organelles. P150 did not associate with lysosome and mitochondria according to the observation under Confocal microscope, but might be colocalized with endoplasmic reticulum (ER) under fluorescence microscope. The P150 of WSSV may participate in virus assembly.