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水產研究(1993年創刊)

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  • 日期: 2019-10-15
水產研究(1993年創刊)-尼羅吳郭魚抗菌胜肽β-防禦素之基因選殖與分子特性研究
主題: 尼羅吳郭魚抗菌胜肽β-防禦素之基因選殖與分子特性研究
摘要(中):
本研究利用表現序列標誌 (EST) 篩選法自尼羅吳郭魚 (Oreochromis niloticus) 選殖到一個屬於 β-防禦素的抗菌胜肽基因,並命名為 OnBD 基因。完整的 OnBD 基因可轉譯出由 66 個胺基酸組成的 prepeptide,此 prepeptide 又可分為兩個功能單位:N-端的 24 個胺基酸組成訊息胜肽,C-端的 42 個胺基酸則為成熟的 OnBD 抗菌胜肽。OnBD 抗菌胜肽的分子組成具有魚類 β-防禦素的家族特徵,其中包含會影響分子構型的 6 個半胱胺酸,此外,OnBD為同時具有親水端與疏水端的兩性分子,帶正電 (淨電荷為+3),不超過 100 個胺基酸序列,這些分子特徵均與抗菌胜肽相符。本研究以即時定量 RT-PCR 方法,分析尼羅吳郭魚 OnBD 之基因表現組織分布,在各組織 (包括血液、鰓、頭腎、腸道、肝、肌肉、皮膚及脾臟) 均可檢測到 OnBD 所表現的 mRNA,其中又以皮膚的表現程度最高。而化學合成的 OnBD 胜肽則用來針對 8 種常見水產病原菌進行最小抑制濃度 (MIC) 的測定,結果顯示 OnBD 對吳郭魚主要的病原菌 Edwardsiella tarda 及 Streptococcus iniae 均有較佳之抑制活性 (其 MIC 可達 4 µM)。綜合以上分子特性與生理活性,可推斷抗菌胜肽 OnBD 在尼羅吳郭魚的先天免疫系統中扮演重要的功能,並後續研究將可應用於水產細菌性疾病之防治。
出版日期: 2015/6/30
標題title(英): Molecular Cloning and Characterization of a β-defensin Antimicrobial Peptide Gene in Nile Tilapia (Oreochromis niloticus)
作者: 陳力豪‧胡鄴方‧朱惠真‧吳嘉哲‧張錦宜
卷別: 23
期別: 1
頁碼: 35-47
作者 auther(英): Li-Hao Chen, Yeh-Fang Hu, Huei-Jen Ju, Chia-Che Wu and Chin-I Chang
摘要abstract(英): An antimicrobial peptide gene named OnBD, which belongs to the β-defensin family, was cloned in Nile tilapia (Oreochromis niloticus) using the expressed sequence tag (EST) cloning method. The putative 66-residue OnBD prepeptide consists of two domains: an N-terminal 24-residue signal domain (signal peptide) and a 42-residue C-terminal antimicrobial domain (mature peptide). Annotation of the antimicrobial domain architecture showed that the OnBD peptide contains the signature motif and six conserved cysteines, a characteristic similar to that of β-defensins in other fish. The OnBD mature peptide shared other common features of antimicrobial peptides, such as being amphipathic and cationic (+3 charge) and having a small molecule (less than 100 a.a.). We also used real-time RT-PCR to investigate the expression pattern of the OnBD gene. The results showed that mRNA of OnBD could be detected in various tissues including the blood, gills, head kidney, intestine, liver, muscle, skin and spleen of Nile tilapia. Quantitative analysis revealed that skin had the highest expression level. Minimum inhibitory concentrations (MICs) of the chemically synthesized OnBD were examined against eight different aquatic bacterial pathogens and shown to exhibit potent antimicrobial activities to all of them, especially to Edwardsiella tarda and Streptococcus iniae, the two most important pathogens to tilapia, with the MIC reaching as low as 4 µM. The molecular characteristics and bioactivities of OnBD suggested that this antimicrobial peptide may play an important role in the innate immunity of Nile tilapia and that it has potential applications in the treatment of bacterial infections.
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