pGreen-S: a clone vector bearing absence of enhanced green fluorescent protein for screening recombinants.
pGreen-S: a clone vector bearing absence of enhanced green fluorescent protein for screening recombinants.
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DOI：
10.1016/j.ab.2009.02.007
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发表时间：
2009-05
影响因子：
2.9
通讯作者：
Jinbao Tang;Shujuan Liang;Jin-bao Zhang;Zhiqin Gao;Suhua Zhang
中科院分区：
文献类型：
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作者：
Jinbao Tang;Shujuan Liang;Jin-bao Zhang;Zhiqin Gao;Suhua Zhang
将增强型绿色荧光蛋白基因插入到pUC18的XbaI酶切位点，构建了细菌克隆载体pGreen-S。在大肠杆菌DH5α中表达时，菌落在日光下呈苦艾绿色，在长波紫外光下呈强荧光绿色。利用pGreen-S载体，根据绿色荧光蛋白缺失导致重组菌落中绿色荧光的丧失，选择定向插入。EGFP报告系统不同于LacZ的传统互补，使筛选重组子更简单、更便宜、更有效。
The bacterial cloning vector, pGreen–S, was constructed by inserting the enhanced green fluorescent protein (EGFP) gene at the XbaI restriction site of pUC18 plasmid. When expressed in Escherichia coli DH5α, it produced colonies that were an absinthe green color under daylight and strongly fluorescent green under longwave ultraviolet light. The pGreen–S vector was used to select for directional insert based on the loss of green fluorescence in recombinant colonies that was caused by the absence of EGFP. The EGFP reporter system differs from the conventional complementation of lacZ, making screening recombinants simpler, less expensive, and more effective.