碩士 / 國立屏東科技大學 / 生物科技系所 / 105 / Viral pathogens is one of major cause of disease outbreaks in aquaculture. It often causes a large amount of death and severe economic loss in aquaculture. Myxovirus-resistance (Mx), which induced by type 1 Interferon (IFN), is one of innate immune members and acts important reole in antiviral function. In the present study, medaka (Oryzias latipes) was used as an animal model to study the immune response and antivial function of Mx genes for NNV infection. In this study, two Mx genes, Mx and Mx-like genes, were cloned from medaka. The full-length of Mx gene is 2286 bp consist of 88 bp of 5’ UTR (5’ untranslated region), 323 bp of 3’ UTR and 1875 bp coding region, which putative encode 624 amino acids. The coding region of Mx-like gene comprises 1479 bp which putative encoded 492 amino acids. Amino acid sequence aligment revraled that Mx comprised GTPase domain in N terminus, GTPase effector domain in C terminus and Middle domain; however Mx-like lack GTPase effector domain in C terminus. Expressional profile of Mx genes in diverse developmental stage and tissue distribution were determined by real- time PCR. Results showed that the expressions of Mx and Mx-like were increased accompanied with embryonic development. Mx gene was aboundant expressed in kidney and spleen, and higher expression of Mx-like gene in spleen and intestine. Mx and Mx-like genes were induced in diverse tissues of medaka by NNV infedction suggesting their antiniral function in innate immune system. In vitro, Mx and Mx-like genes were induced respectively in medaka OLHE-131 cells after IFN-a and IFN-d transfection suggesting the expression of Mx and Mx-like were regulated by IFN-a and IFN-d. Functional assay showed that OLHE-131 cells transfected with Mx rxpressing plasmid suppress NNV-induced apoptosis, whereas Mx-like without anti-apoptosis function. This result suggested the Mx and Mx-like may play distinct finction in immune system against NNV infection. The results presented in this study can be used as a platform to screen antiviral substances based on the induction of Mx expression. It is helpful to accelerate the development of biocontrol approaches for viral-induced disease problems in aquaculture.
| Identifer | oai:union.ndltd.org:TW/105NPUS5111004 |
| Date | January 2017 |
| Creators | Lu, Jin-YI, 呂勁逸 |
| Contributors | Hu, Shao-Yang, Hong, Yong-Han, 胡紹揚, 洪永瀚 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 69 |
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