Hossain M S, Khadijah Siti, Kwang Jimmy
Animal Health Biotechnology, Temasek Life Sciences Laboratory, The National University of Singapore, Singapore 117604, Singapore.
Virology. 2004 Jul 20;325(1):106-15. doi: 10.1016/j.virol.2004.05.005.
Open reading frame 89 (ORF89) is one of the three genes that are believed to be involved in the latent infection of white spot syndrome virus (WSSV). Here, we report the structure and functional characterization of ORF89. cDNA sequencing, 5' RLM-RACE, and 3' RLM-RACE showed that ORF89 gene is transcribed into an unspliced mRNA of 4436 nucleotides, which is predicted to encode a protein of 1437 amino acids. ORF89 expressed an approximately 165-kDa protein in Sf9 cells that localized in the nucleus. Amino acids 678-683 were found to be essential for nuclear localization. Cotransfection assays demonstrated that ORF89 protein repressed its own promoter as well as those of a protein kinase and the thymidine-thymidylate kinase genes of WSSV. SYBR Green real-time PCR indicated that the repression occurred at the transcriptional level.
开放阅读框89（ORF89）是被认为参与白斑综合征病毒（WSSV）潜伏感染的三个基因之一。在此，我们报告了ORF89的结构和功能特征。cDNA测序、5' RLM-RACE和3' RLM-RACE表明，ORF89基因转录成一个4436个核苷酸的未剪接mRNA，预计编码一个1437个氨基酸的蛋白质。ORF89在Sf9细胞中表达了一种约165 kDa的蛋白质，该蛋白质定位于细胞核。发现氨基酸678 - 683对于核定位至关重要。共转染实验表明，ORF89蛋白抑制其自身启动子以及WSSV的一个蛋白激酶和胸苷 - 胸苷酸激酶基因的启动子。SYBR Green实时PCR表明这种抑制发生在转录水平。
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