| 绿色荧光蛋白在大肠杆菌中的克隆及高效表达 |
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| 引用本文： | 陆惠萍,周凤娟.绿色荧光蛋白在大肠杆菌中的克隆及高效表达[J].第二军医大学学报,1997,18(5):406-408. |
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| 作者姓名： | 陆惠萍 周凤娟 |
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| 作者单位： | 第二军医大学基础医学部分子遗传学教研室 |
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| 摘 要： | 在大肠杆菌中克隆及高效表达绿色荧光蛋白基因。方法；ＰＣＲ法克隆ＧＦＰ－Ｓ６５ＴｃＤＮＡ并改造其两端的限制性内切酶位点，构建重组原核表达载体ｐＲＳＥＴ－ＧＦＰＳ６５Ｔ。结果；在ＩＰＴＧ诱导条件下，ＧＦＰ－Ｓ６５Ｔ的表达量占细菌蛋白的 上。细菌培养物及其超声上清在长波紫外线的照射下，发出明亮的绿色荧光。
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| 关 键 词： | 基因表达 大肠杆菌 绿色荧光蛋白 克隆 |
Cloning and expression of green fluorescent protein in E.coli |
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| Lu Huiping,Zhou Fengjuan,Sun Shuhan.Cloning and expression of green fluorescent protein in E.coli[J].Academic Journal of Second Military Medical University,1997,18(5):406-408. |
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| Authors: | Lu Huiping Zhou Fengjuan Sun Shuhan |
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| Abstract: | Objective: To clone and express green fluorescent protein S65T in E.coli. Methods: GFP S65T cDNA fragment was cloned with Nde Results: Induced by IPTG, GFP constituted more than 15% of total bacterial proteins. Induced E.coli and its ultrasonication supernatant produced strong green fluorescence when excited by long wave ultraviolet(UV) light. Conclusion: GFP S65T can be used as a report gene in prokaryotes, and its expression can be easily detected by long wave UV. |
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| Keywords: | protein green fluorescent gene expression genes reportorial |
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