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首页> 美国卫生研究院文献>Applied and Environmental Microbiology >Expression of the ggpS Gene Involved in Osmolyte Synthesis in the Marine Cyanobacterium Synechococcus sp. Strain PCC 7002 Revealed Regulatory Differences between This Strain and the Freshwater Strain Synechocystis sp. Strain PCC 6803
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Expression of the ggpS Gene Involved in Osmolyte Synthesis in the Marine Cyanobacterium Synechococcus sp. Strain PCC 7002 Revealed Regulatory Differences between This Strain and the Freshwater Strain Synechocystis sp. Strain PCC 6803

机译：ggpS基因的表达涉及海洋蓝藻Syechococcus sp。中的渗透液合成。菌株PCC 7002此菌株与淡水菌株Synechocystis sp。之间的显示调控差异。应变PCC 6803

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Synthesis of the osmolyte glucosylglycerol (GG) in the marine cyanobacterium Synechococcus sp. strain PCC 7002 was characterized. The ggpS gene, which encodes the key enzyme (GG-phosphate synthase [GgpS]) in GG biosynthesis, was cloned by using PCR. A 2,030-bp DNA sequence which contained one open reading frame (ORF) was obtained. The protein deduced from this ORF exhibited 85% similarity to the GgpS of the freshwater cyanobacterium Synechocystis sp. strain PCC 6803. The function of the protein was confirmed by generating a ggpS null mutant, which was not able to synthesize GG and thus exhibited a salt-sensitive phenotype. Expression of the ggpS gene was analyzed in salt-shocked cells by performing Northern blot and immunoblot experiments. While almost no expression was detected in cells grown in low-salt medium, immediately after a salt shock the amounts of ggpS mRNA and GgpS protein increased up to 100-fold. The finding that salt-induced expression occurred was confirmed by measuring enzyme activities, which were negligible in control cells but clearly higher in salt-treated Synechococcus sp. cells. The salt-induced increase in GgpS activity could be inhibited by adding chloramphenicol, while in protein extracts of the freshwater cyanobacterium Synechocystis sp. strain PCC 6803 a constitutive, high level of enzyme activity that was not affected by chloramphenicol was found. A comparison of GG accumulation in the two cyanobacteria revealed that in the marine strain osmolyte synthesis seemed to be regulated mainly by transcriptional control, whereas in the freshwater strain control seemed to be predominantly posttranslational.
机译：海洋蓝藻Syechococcus sp。中渗透性葡萄糖基甘油（GG）的合成。表征了PCC 7002菌株。使用PCR克隆了编码GG生物合成中的关键酶（GG-磷酸合酶[GgpS]）的ggpS基因。获得了一个包含一个开放阅读框（ORF）的2030 bp DNA序列。由该ORF推导的蛋白质与淡水蓝藻集胞藻Syechocystis sp。的GgpS具有85％的相似性。通过产生ggpS无效突变体证实了蛋白质的功能，该ggpS无效突变体不能合成GG，因此表现出对盐敏感的表型。通过进行Northern印迹和免疫印迹实验，分析了盐冲击细胞中ggpS基因的表达。虽然在低盐培养基中生长的细胞中几乎未检测到表达，但在盐冲击后，ggpS mRNA和GgpS蛋白的量立即增加了100倍。通过测量酶活性可以证实盐诱导表达的发现，该酶活性在对照细胞中可以忽略不计，但在盐处理的Synechococcus sp。中明显更高。细胞。盐诱导的GgpS活性增加可通过添加氯霉素来抑制，而在淡水蓝藻集胞藻属（Synechocystis sp。）的蛋白质提取物中。发现PCC 6803菌株是一种不受氯霉素影响的高水平组成型酶活性。两种蓝藻中GG积累的比较表明，在海洋菌株中，渗透液的合成似乎主要受转录控制，而在淡水菌株中，调控似乎主要在翻译后。

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