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題名 | 創傷弧菌Vibrio vulnificus藍色螢光蛋白產光原因之探究=Study of Fluorescence Production on Blue Fluorescent Protein from Vibrio Vulnificus |
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作者 | 蘇哲弘; 陳連輝; 謝桂鈺; 張淑玉; | 書刊名 | 嘉南學報 |
卷期 | 26 2000.11[民89.11] |
頁次 | 頁130-140 |
分類號 | 414.83 |
關鍵詞 | 創傷弧菌; 藍色螢光蛋白質; 短鏈去氫氧化還原酶; Vibrio vulnificus; Blue fluorescent protein; Short-chain dehydrogenase/reductase; |
語文 | 中文(Chinese) |
中文摘要 | 創傷弧菌Vibrio vulnificus CKM-1的藍色螢光蛋白基因(bfp),可譯出239個氧基酸序列,將bfp in frame接入pET21b表現載體,轉形進入大腸桿面BL21,經由長波UV燈(366nm)照射會發出顯著的藍色螢光;透過多重氨基酸序列比對,得知藍色螢光蛋白質(BEP)應歸類於短鏈去氫氧化還原?(short-chain dehydrogenase/reductase)(SDR)family中之一員,而且BEF亦具有SDR蛋白質特有的絕對保留性殘基;除此之外,不同來源的SDR蛋白質利用X射線結晶圖也已證實其立體構形的型式具有高度的相似性。經由人類cDNA基因庫將五種SDR基因選殖出來,分別以pET21b為載體,轉形進入大腸桿菌BL21,發現這些與BFP具有同源性的SDR基因產物並不具產光的特性,初步推論BFP可以直接吸收菌體外的輻射性光能(UV),並將能量轉移而釋出螢光的產光特性,可能不單是取決於分子的三度空間結構。 |
英文摘要 | The blue fluorescent protein gene (bfp) from Vibrio vulnificus CKM-1 encoded 239 amino acids. The bfp was in frame constructed into pET21b and expressed in Escherichia coli BL21, the transformant exhibited distinct blue fluorescence after illuminating with a long wave ultraviolet (UV) source in the dark. The nucleotide sequence of bfp gene and its deduced amino acid sequence showed significant homology to those of the short-chain dehydrogenase / reductase (SDR) family proteins from various organisms. Likewise, some functionally important residues in SDR are strictly conserved in BFP. Conformational patterns of SDR enzymes are highly similar except for variations in the C-terminal parts when they are established by X-ray crystallography. Five SDR protein genes from human were cloned and transformed to E. coli BL21. the fluorescence was not seen in transformants. We suggested that the blue fluorescence production of BFP is probably a consequence of energy transfer from UV to the BFP by radiative process and this trait may not be established by conformational patterns of SDR protein. |
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