Detection of rhythmic bioluminescence from luciferase reporters in cyanobacteria.
Detection of rhythmic bioluminescence from luciferase reporters in cyanobacteria.
复制标题
检测蓝藻中荧光素酶报告基因的节律性生物发光。
DOI：
10.1007/978-1-59745-257-1_8
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发表时间：
2007
期刊：
影响因子：
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通讯作者：
Golden,SusanS
中科院分区：
文献类型：
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作者：
Mackey,ShannonR;Ditty,JaynaL;Clerico,EugeniaM;Golden,SusanS
单细胞蓝藻长聚球菌pcc 7942是研究原核生物昼夜节律的模式生物。虽然。长尾藤没有明显的昼夜节律行为，其基因表达受昼夜节律控制；因此，一种“行为”是通过将蓝藻启动子连接到细菌荧光素酶基因或荧光素酶基因来创建报告融合体，其活性可以很容易地通过生物发光来监测。我们的实验室已经创建了许多载体来将荧光素酶报告基因引入到这些细胞中。elongatuschromosome。检测荧光素酶融合产生光的方法和设备的选择提供了一种高通量、自动化突变筛选的手段，以及在同一细胞培养中测试两个启动子融合的节律。
The unicellular cyanobacteriumSynechococcus elongatusPCC 7942 is the model organism for studying prokaryotic circadian rhythms. AlthoughS. elongatusdoes not display an easily measurable overt circadian behavior, its gene expression is under circadian control; hence, a “behavior” is created by linking a cyanobacterial promoter to either the bacterialluxABor fireflylucluciferase genes to create reporter fusions whose activity can be easily monitored by bioluminescence. Numerous vectors have been created in our lab for introducing luciferase reporter genes into theS. elongatuschromosome. A choice of methods and equipment to detect light production from the luciferase fusions provides a means for high-throughput, automated mutant screens as well as testing rhythms from two promoter fusions within the same cell culture.