細胞自噬是細胞內一種重要的機制，主要透過形成自噬小體和自噬溶酶體，降解細胞質中非必需蛋白質，減少外來物質與壓力之干擾，進而促進細胞正常之生長與發展。LC3蛋白對於自噬小體形成過程中扮演關鍵角色，並被廣泛視為是否產生細胞自噬之重要標記。點帶石斑魚過去相關研究，並無選殖及開發該物種之LC3作為生物標記。本研究嘗試選殖點帶石斑魚的LC3基因並針對該基因特性進行探討。該基因全長1440 bp可轉譯 126氨基酸長之蛋白。此外，LC3基因可在各組織表現。為了解 LC3 於石斑魚生長期表現情形，研究中觀察其石斑魚卵孵化後不同時間點之基因表現，結果顯示基因能在不同批的魚苗有相同的趨勢。經Hank’s solution培養及rapamycin處理GF-1細胞下，証實點帶石斑LC3會轉換成LC3-II形成自噬小體和自噬溶酶體。再者，無論是細胞亦或是動物實驗，顯示經病毒感染後皆可誘發點帶石斑LC3之表現，進一步經研究証實點帶石斑LC3及Hsp90AB彼此間具有交互作用，未來可深入研究LC3蛋白與 Hsp90AB於病毒感染機制上所扮演之角色為何。

Autophagy is a conserve and important cellular mechanism to sustain cell survival during stress, promote healthy growth and development by degrade and recycle unwanted cytosolic proteins and organelles in autophagosome and autolysosome. LC3, involved in autophagosome formation is widely used as marker. However, currently there is no orange-spotted grouper (Epinephelus coioides) LC3 cloned. In this study, a grouper’s LC3 (osgLC3) mRNA had been cloned. Its size is 1440 bp DNA sequence and can be translated to 126 amino acid long proteins. Besides, expression level of osgLC3 found constitutively expressed in all tissues. For the investigation of osgLC3 expression level during grouper’s larvae development, sample from different days of post-hatchery were examined found similar pattern of expression from 3 different batches. Moreover, treatments of Hank’s solution and rapamycin on GF 1 cell line, the osgLC3 was able to form osgLC3-II and synthesizes autophagosome and autolysosome. Furthermore, it has found that autophagy was induced during virus infection, not only in vitro, but also in vivo experiments. Interesting discovery of protein-protein interaction between osgLC3 and Hsp90AB allows future study on their roles and mechanisms during NNV infection in orange-spotted grouper.

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