參賽序號：24
海報主題
探討麩胺酸丙酮酸轉胺酶(GPT)參與WSSV誘發代謝改變現象之角色
系級
生物科技與產業科學系
指導老師及參賽學生
指導老師：王涵青
參賽學生：鄭淑文
構想說明
White spot syndrome virus (WSSV) causes huge economic losses in shrimp farming in Asia. The molecular mechanisms behind the WSSV pathogenesis are a target for developing a method to control the disease. WSSV triggers glutaminolysis in shrimp to produce biomolecules and energy to support replication. Our objective was to characterize an enzyme called Glutamic-Pyruvic Transaminase (GPT), involved in glutamate metabolism, in WSSV-infected shrimp. In humans, GPT was in cytosolic and mitochondrial forms, namely GPT1 and GPT2, respectively. The sequence of shrimp GPT1 (LvGPT1) was constructed using primer sets designed based on our in-house transcriptomic database. Cellular localization of LvGPT1 was studied with immunofluorescence (IF); LvGPT1 was localized in the cytosol, similar to human GPT1. dsRNA silencing of LvGPT1 significantly reduced viral gene expression and genome replication. Overall, we inferred that LvGPT1 had a pivotal role in virus replication. In future studies, we will examine whether the virus replication can be rescued after LvGPT1 silencing by replenishing a downstream metabolite of GPT1. Second, we will use Yeast 2-hybrid to investigate interactions between WSSV viral protein and LvGPT1, as this might provide an insight regarding how WSSV proteins regulate LvGPT1. This study demonstrated the importance of LvGPT1 in WSSV replication and provided a potential selective marker for breeding shrimp with anti-virus ability.
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