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首页> 外文期刊>Journal of bacteriology >Cloning, Sequencing, and Phenotypic Characterization of the rpoS Gene from Pseudomonas putida KT2440
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Cloning, Sequencing, and Phenotypic Characterization of the rpoS Gene from Pseudomonas putida KT2440

机译：恶臭假单胞菌KT2440 rpoS基因的克隆，测序及表型鉴定

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A gene homologous to the rpoS gene of Escherichia coli was cloned from a Pseudomonas putida KT2440 gene bank by complementation of the rpoS-deficient strainE. coli ZK918. The rpoS gene of P. putida complemented the acid sensitivity and catalase deficiency of the rpoS mutant of E. coli and stimulated expression of the RpoS-controlled promoter,bolAp 1. The gene was sequenced and found to be highly similar to the rpoS genes of other gram-negative bacteria. Like in other gram-negative bacteria, a homolog of thenlpD gene was found upstream to the rpoS gene. A transcriptional fusion of the promoter of the P. putida rpoS gene to the luxAB genes from Vibrio harveyi was constructed and used as an inactivated allele ofrpoS for gene replacement of the wild-type copy in the chromosome of P. putida. The resultantrpoS mutant of P. putida, C1R1, showed reduced survival of carbon starvation and reduced cross-protection against other types of stress in cells starved for carbon, in particular after a challenge with ethanol. Survival in soil amended with m-methylbenzoate was also reduced in the mutant strain P. putida C1R1. The RpoS protein ofP. putida controls the expression of more than 50 peptides, which are normally expressed in cells after a short period of carbon starvation.
机译：通过补充 rpoS 从恶臭假单胞菌 KT2440基因库中克隆了与大肠杆菌的 rpoS 基因同源的基因。 em>-缺陷株 E。大肠菌 ZK918。 P的 rpoS 基因。 putida 补充了 E的 rpoS 突变体的酸敏感性和过氧化氢酶缺陷。大肠杆菌和受RpoS调控的启动子 bolAp 1 的刺激表达。对该基因进行了测序，发现与其他革兰氏阴性细菌的 rpoS 基因高度相似。与其他革兰氏阴性细菌一样，在 rpoS 基因的上游发现了 nlpD 基因的同源物。 P启动子的转录融合。构建了哈维弧菌 luxAB 基因中的putida rpoS 基因，并将其用作 rpoS 的失活等位基因，用于取代在 P染色体上的野生型拷贝。恶臭。产生的 P的 rpoS 突变体。 putida （C1R1）在缺乏碳的细胞中表现出碳饥饿的存活率降低，并且对其他类型压力的交叉保护作用降低，尤其是在受到乙醇攻击后。在 P 突变菌株中，用 m -甲基苯甲酸甲酯改良的土壤中的存活率也降低了。恶臭 C1R1。 P的RpoS蛋白。 putida 控制着50多种肽的表达，这些肽通常在短时间的碳饥饿后在细胞中表达。

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