Research on Nervous Necrosis Virus Controlling GB3 Cell Gene Expression
| Autor: | Chung-Hsiang Yuan, 袁崇翔 |
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| Rok vydání: | 2007 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 95 Following infection, viruses use different mechanisms to control host cell’s genes expression. Some of the host genes may be activated or suppressed. And its makes the host cell to be an appropriate place for virus to replicate and assemble. Recently, viral nervous necrosis disease causes a great damage to grouper aquaculture in Taiwan. The pathogen of this disease has been identified as nervous necrosis virus (NNV). In order to understand the infection characteristics of this virus, first we established a phage display cDNA library of virus’s host cell, grouper brain cell (GB3), and used immunostaining method to screen the differentially-expressed genes in response to NNV infection. These screening resulted in the identification of 46 differentially-expressed genes. There were 18 genes been activated, 27 been suppressed and 1 was constantly expressing gene. Analyzing and comparing these genes sequences with NCBI genetic database, it was observed that some of the up-regulated relate to protein translation, like translation elongation factor 1-delta 、ribosomal protein S18、ribosomal protein S10 and ribosomal protein LP0. There has also immune-relate gene: natural killer cell enhancement factor, which may activate the host cell’s immuno-system. Besides, ADP/ATP translocase and Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) genes which relate to energy production have also been found to be up-regulated. A many down-regulated genes, endozepine and S100-like calcium binding protein, which relate to neuro-transmittance and abundant in brain cell, emerge frequently. The nucleic acid-metabolic gene, which related to cell-cycle, nucleoside diphosphate kinase and some protein translation-related genes such as 60S ribosomal protein L27 and ribosomal protein L23 were down- regulated. From this result, we assume that the selected protein translation-related genes may be utilized by the virus to produce viral proteins. The results of RT-PCR, besides confirming the immunostaining screening, showed the differentially expressed genes at transcription level. This experiment provides a new approach to study the difference in host gene expression at transcription and translation levels after virus infection. Taken together, these results revealed the up- and down-regulated genes in response to NNV infection, and this information could be helpful to proceed further to understand the molecular mechanisms behind the NNV-host interaction and to develop control measures against NNV. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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