博士 / 國立臺灣海洋大學 / 海洋生物研究所 / 103 / White spot syndrome virus (WSSV) is a large enveloped DNA virus, and it causes a serious disease that has led to severe mortalities of cultured shrimps in many countries. To determine the mechanism of virus entry into the cell and to establish an antiviral strategy, the cell receptor for virus entry and receptor binding protein should be determined. In this study, a surface protein, named glucose transporter 1 (Glut1), which could interact with WSSV envelope protein VP53A was identified. Sequence analysis revealed that Glut1 closely related a member of a large superfamily of transporters that function to transport sugar, and Glut1 was confirmed to have the ability of transporting glucose. This Glut1 gene can also be found in other shrimp species. Glut1 protein was 68 kDa moelculare weight, and was constitutive in almost all organs. The interaction between Glut1 and some other WSSV envelope proteins in the infectome structure was verified by far western blot and His pull down assay. In vitro and in vivo neutralization using recombinant partial Glut1 revealed that the large extracellular portion of Glut1 could delay WSSV infection. Also, the mortality of the shrimps which were knocked-down the Glut1 gene by treated with dsRNA before WSSV challenged showed decreased. These results indeed provide a direction to develop efficient antiviral strategies or therapeutic methods by using Glut1.
Besides, Glut1 gene was discovered to be suppressed in all tissues after WSSV infection, except midgut. Yeast two-hybrid was performed using Glut1 as the bait to screen the proteins that can interact with Glut1, and the result showed that Glut1 could bind with WSSV 183/184 proteins very strongly. The data indicated that WSSV 183/184 probably affect the expression and function of Glut1 after WSSV infection. These findings will have important implications for our understanding of shrimp pathogenicity in WSSV infection.
| Identifer | oai:union.ndltd.org:TW/103NTOU5270006 |
| Date | January 2014 |
| Creators | Huang, Huai-Ting, 黃懷葶 |
| Contributors | Chen, Li-Li, 陳歷歷 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 75 |
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