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Detection of the isiA Gene across Cyanobacterial Strains: Potential for Probing Iron Deficiency

机译:跨蓝细菌菌株isiA基因的检测:探测铁缺乏症的潜力

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摘要

The use of isiA expression to monitor the iron status of cyanobacteria was investigated. Studies of laboratory cultures of the cyanobacterium Synechocystis sp. strain PCC 6803 showed that isiA expression is dependent on the organism's response to iron deficiency; isiA expression starts as soon as a decline in the rate of growth begins. isiA expression is switched on at concentrations of iron citrate of less than 0.7 μM. A PCR method was developed for the specific amplification of the iron-regulated isiA gene from a variety of cyanobacteria. After we developed degenerate primers, 15 new internal isiA fragments (840 bp) were amplified, cloned, and sequenced from strains obtained from algal collections, from new isolates, and from enriched field samples. Furthermore, isiA expression could be detected by means of reverse transcription-PCR when enriched field samples were exposed to restricted iron availability. These results imply that determining the level of iron-regulated isiA expression can serve to indicate iron deficiency in cyanobacterial samples of differing origins from the field.
机译:研究了使用isiA表达监测蓝细菌的铁状态。蓝藻Synechocystis sp。的实验室培养物的研究。 PCC 6803菌株显示isiA表达取决于生物体对铁缺乏的反应。一旦增长率开始下降,isiA表达就会开始。在小于0.7μM的柠檬酸铁浓度下开启isiA表达。开发了一种PCR方法,用于从多种蓝细菌中特异性扩增铁调节的isiA基因。在我们开发了简并引物之后,从藻类收集物,新分离物和富集野外样品中提取了15个新的内部isaA片段(840 bp),进行了克隆和测序。此外,当富集的野外样品暴露于有限的铁利用率下时,可通过逆转录PCR检测isiA表达。这些结果暗示确定铁调节的isiA表达的水平可以用来指示来自不同领域的蓝细菌样品中的铁缺乏。

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