A promoter-trap vector for clock-controlled genes in the cyanobacterium Synechocystis sp PCC 6803
A promoter-trap vector for clock-controlled genes in the cyanobacterium Synechocystis sp PCC 6803
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DOI：
10.1016/s0167-7012(01)00376-1
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发表时间：
2002-05-01
影响因子：
2.2
通讯作者：
Ishiura, M
中科院分区：
文献类型：
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作者：
Aoki, S;Kondo, T;Ishiura, M
我们构建了一个启动子捕获载体pPT 6803 -1，以分离蓝藻集胞藻菌株PCC 6803中的昼夜节律钟控制的启动子。该载体含有来自哈氏弧菌的无启动子荧光素酶基因组（luxAB），其靶向集胞藻基因组的特定位点作为基因表达的报告基因。通过将基因组DNA片段引入luxAB上游以转化集胞藻细胞，在pPT 6803 -1中构建文库。在大约10，000个集胞藻转化体中，至少55个（#1-55）在连续光照下显示生物发光的昼夜节律。克隆#19、#22和#26表现出彼此明显不同的生物发光波形。缺失分析和引物延伸实验绘制了clpP的启动子。slr 1634和rbpP基因，它们分别来自#19、#22和#26，负责生物发光。(C)2002 Elsevier Science B. V.保留所有权利。
We constructed a promoter-trap vector pPT6803-1 to isolate circadian clock-controlled promoters in the cyanobacterium Synechocystis sp. strain PCC 6803. The vector contains a promoterless luciferase gene set (luxAB) from Vibrio harveyi that is targeted to a specific site of the Synechocystis genome as a reporter for gene expression. A library was constructed in pPT6803-1 by introducing the genomic DNA fragments upstream of luxAB to transform Synechocystis cells. Of approximately 10,000 Synechocystis transformants, at least 55 (#1-55) showed circadian rhythms of bioluminescence under continuous illumination. Clones #19, #22, and #26 exhibited obviously different waveforms of bioluminescence from each other. Deletion analysis and primer extension experiments mapped the promoters for the clpP. slr1634, and rbpP genes that are responsible for biolumineseence from #19, #22, and #26, respectively. (C) 2002 Elsevier Science B.V. All rights reserved.