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標題: Preliminary study on the regulation of cecropin gene expression in Armigeres subalbatus
作者: 蘇永倫
Yung-Luen Su
關鍵字: cecropin;白腹叢蚊;抗微生物胜肽
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本論文利用白腹叢蚊(Armigeres subalbatus)進行抗微生物胜肽cecropin基因表現調控的研究,選殖總計長度1939 bp之白腹叢蚊cecropin A2轉錄單位前之五端序列,並將該1939 bp五端序列片段截切,利用pGL4.10[luc2]報導基因載體構築成1939 CecA2-pGL4、1609 CecA2-pGL4與1184 CecA2-pGL4質體。將這些質體轉染於C6/36細胞中並以大腸桿菌(Escherichia coli)與金黃色葡萄球菌(Staphylococcus aureus)進行luciferase基因表現誘導作用。在大腸桿菌的誘導處理組,以1609 CecA2-pGL4質體的RNA偵測量達對照組的4.948倍為最高;而於金黃色葡萄球菌的誘導處理組,各質體表現量無顯著差異。此外,本論文亦嘗試建構白腹叢蚊體內報導基因表現系統,用以探討cecropin基因五端轉錄作用調節區的誘導表現。蚊體試驗先測試注射上述質體,或質體轉染試劑cellfectin II,結果顯示受測之雌蚊存活率為97%與83%,且利用即時定量聚合酶連鎖反應 (real-time PCR)檢測引子增幅效能均達90%以上。同時1939 CecA2-pGL4質體在注射進雌蚊體內後可偵測得報導基因luciferase表現之RNA,顯示白腹叢蚊蚊體可以進行外源基因調控區研究之應用。在蚊體注射1939 CecA2-pGL4、1609 CecA2-pGL4與1184 CecA2-pGL4質體後,再利用大腸桿菌進行誘導作用,初步結果證實cecropin A2五端各片段區域具有基因轉錄作用調控現象,其表現模式與C6/36細胞結果相似。
其他識別: U0005-0302201516503400
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