中文摘要

　　本研究主要探討創傷弧菌藍色螢光蛋白基因(bfpvv)在大腸桿菌Escherichia coli XL1-Blue巨大原生質體表現之可行性，並確立以螢光顯微鏡及數位照相定量菌體內創傷弧菌藍色螢光蛋白螢光強度之方法。
　　實驗發現含bfpvv基因之大腸桿菌原生質體可以放出螢光，具有細胞功能性，但因生產螢光蛋白造成原生質體死亡率較原來宿主大腸桿菌原生質體高。定量巨大原生質體及正常菌體之螢光強度，發現巨大原生質體螢光強度高於正常菌體且螢光維持時間較正常菌體長。此意味著基因組換菌在原生質體化及巨大化過程中質體不但沒有脫落，且產物之螢光蛋白濃度亦高於正常菌體。利用此結果，本研究之大腸桿菌巨大原生質體在生物取像系統具發展潛力。

Abstract

The expression of Vibrio vulnificus gene encoding blue fluorescence protein (bfpvv) by Escherichia coli XL1-Blue giant protoplast was investigated for determining the method for quantifying the intensity of fluorescence emitted by the blue fluorescence protein by means of fluorescence microscopy and digital image analysis.
The result indicated that fluorescence could be emitted by the giant protoplast containing bfpvv gene, which showed complete cell function. However, the viability of the bfpvv gene-carrying protoplast was lower than the bfpvv gene-free protoplast because the expression of bfpvv gene posed a metabolic burden on the cells. As compared with normal cells contain bfpvv gene, the fluorescence emitted by the giant protoplast containing bfpvv gene was more intense and permanent. It was shown that the plasmid containing bfpvv gene was maintained in the cells during the formation of giant protoplast produced more fluorescence protein than the normal cells. The giant protoplast of E.coli is potential for bioimaging system.

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