本研究在於探討創傷弧菌藍色螢光蛋白基因 (bfp) 與經由DNA變異法所得突變基因 (bfp-d7) 在革蘭氏陰性菌Escherichia coli XL1-Blue與Vibrio vulnificus CKM-1巨大原生質體之螢光表現，並利用巨大原生質體評估重組基因啟動子強度之方法。

實驗中定義兩種螢光值，分別為比螢光值與總螢光值，比螢光值為去單位化Image-Pro Plus所分析出螢光色階之數據，而總螢光值為比螢光值乘上光學密度(Optical Density 600)，可正規化比螢光值，以利起動子效率的估算。啟動子效率偵測方面，共有三種變數，每種變數有兩個參數，分別為菌株 (E.coli XL1-Blue、V.vulnificus CKM-1)、螢光蛋白 (BFP、BFP-D7) 與溫度 (30°C、25°C)，搭配起來有8種變化，並根據總螢光值對時間之曲線的最高點到最低點範圍，以線性趨勢線逼近，得趨勢線斜率，代表啟動子效率。比較以上各種變化之啟動子效率，得到一偵測啟動子強度之最佳系統為利用菌株E.coli XL1B在BFP-D7螢光蛋白表現下，於30°C偵測啟動子強度。

The expression of blue fluorescent protein gene (bfp) and mutated gene (bfp-D7) from bfp by random mutagenesis isolated from Vibrio vulnificus CKM-1 in the giant protoplasts of Escherichia coli XL1B and V. vulnificus CKM-1 were investigated, respectively. In addition, we built a system for quantifying the fluorescent intensity of BFP and screened a high efficiency promoter.

We defined two kinds of fluorescence density, one was specific blue density that was de-unit data from Image-Pro Plus’ color depth ; another’s overall blue density that was from specific blue density multiplied optical density. The overall blue density was able to represent specific blue density normalization and evaluated transcription efficiency in promoter. About promoter efficiency evaluated, we had three kinds of variables including two parameters with eight variations, one was bateria including E.coli XL1-Blue and V. vulnificus CKM-1, one was fluorescence protein including BFP and BFP-D7, another was temperature including 30°C and 25°C. According the diagram of overall blue density varied with time, we found the slope of regression line fitting the curve which range was from the highest fluorescence value to the lowest value. The slope represented promoter efficiency. The results showed that the most adaptable system to detect promoter intensity was E.coli XL1-Blue_pUD7 in 30°C.

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