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頁籤選單縮合
題 名 | Mutation Analysis of Pertussis Toxin Promoter=百日咳基因啟動子之突變分析 |
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作 者 | 吳游源; 邱淑君; 盧政雄; | 書刊名 | 微免與感染雜誌 |
卷 期 | 32:3 1999.09[民88.09] |
頁 次 | 頁163-172 |
分類號 | 415.279 |
關鍵詞 | 突變; 百日咳毒素; Pertussis toxin; Mutation; Promoter; |
語 文 | 英文(English) |
中文摘要 | 分析改良表現效率相當弱的百日咳毒素基因啟動子是非細胞性疫苗研製的一個重 要議題。在本實驗中,我們由百日咳菌株 ATCC9340 中製備了百日咳毒素啟動子區域。根據 所獲得的啟動子核酸序列,建構了一系列的突變型啟動子。然後利用體外膠體移轉試驗及體 內β -galactosidase 之活性測定以分析其相對效率。 結果發現, 若將核醣體鍵結序列或 -10 區域序列改換成較強的細菌啟動子序列,可使突變後之啟動子與兩個菌體中蛋白之鍵結 增強,並得到稍高之β -galactosidase 活性(野生型的 1.3 倍)。 若將較遠距之反向或 20- 核酸對之直接重複序列改換成某特定之完全重複序列,不但可增強啟動子與另一高分子 量菌體蛋白之鍵結,同時可表現更高的β -galactosidase 活性( 1.8 倍)。 此項發現對 提高百日咳毒素產率之突變型菌株之建構,提供另一個研發途徑。 |
英文摘要 | The improving of the expression efficiency of a pertussis toxin (PT) promoter was believed to be a critical issue for the production of PT in acellular vaccine development. In this study, we have isolated a PT promoter region from the genome of a pertussis strain ATCC 9340. Based on the promoter sequence, a series of mutant PT promoters have been generated and subjected to in vitro gel shift analysis and in vivo reporter β -galactosidase activity study. As compared with the wild type promoter, the mutation of the ribosome binding sequence or -10 element, to the respective consensus sequence derived from strong bacterial promoters, resulted in an enhancement of its interaction with two cellular proteins, and a slightly higher β -galactosidase activity (1.3 fold). Whereas, the change of either upstream inverse repeats or 20-bp direct repeats to a certain complete repeat significantly promoted the formation of another DNA-protein-complex, and exhibited an 1.8 fold β -galactosidase activity. These findings would have provided a mutation target for making a more efficient PT-production pertussis strain. |
本系統中英文摘要資訊取自各篇刊載內容。