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作 者 ：田丽芳，魏兰珍，王全喜*

期 刊 ：西北植物学报 2013年 7期 页码：1311~1316

关键词：ndhK；启动子；转录调控；

Keywords:ndhK, promoter, transcriptional regulation,

摘 要 ：ndhK是蓝藻NDH-1复合体的1个亚基编码基因，其表达受低浓度CO₂和高光的诱导，对于蓝藻应对低CO₂胁迫起着重要的作用。为进一步阐明ndhK基因的转录调控机制，本研究利用5′-RACE(Rapid Amplification of cDNA Ends,cDNA末端快速扩增)技术鉴定了该基因的转录起始位点，利用生物信息学预测发现ndhK基因含有4个可能的启动子，构建了含增强型黄色荧光蛋白报告基因的启动子探针型载体，并利用蛋白免疫印迹的方法进行检测。结果表明：在集胞藻6803的ndhK基因上游的-374～-274 bp，-438～-374 bp，-604～-543 bp，+1～+52 bp区域具有较高的启动子活性，而-543～-440 bp区域则可能存在1个抑制ndhK基因表达的转录因子。

Abstract:ndhK is a subunit of cyanobacteria NDH-1 complex,playing an important role in responding to low CO₂ stress.Its expression level is induced by low CO₂ and high light.To further understand the mechanism of transcriptional regulation of ndhK with light,we find the TSP(transcription start position) with 5′-RACE(Rapid Amplification of cDNA Ends),forecast that ndhK contain four possible promoter,construct a series of promoter probe vectors containing eYFP (enhanced yellow fluorescent protein) and use western blot to detect them.The results showed that the upstream sequence of ndhK in Synechocystis sp.PCC6803 containing four sequences (-374 to -274，-438 to -374，-604 to -543，+1 to +52) which can promote gene expression and one can inhibit (-543 to -440).

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