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首页> 外文期刊>Journal of bacteriology >Analysis of Pseudomonas putida KT2440 Gene Expression in the Maize Rhizosphere: In Vitro Expression Technology Capture and Identification of Root-Activated Promoters
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Analysis of Pseudomonas putida KT2440 Gene Expression in the Maize Rhizosphere: In Vitro Expression Technology Capture and Identification of Root-Activated Promoters

机译：恶臭假单胞菌KT2440基因在玉米根际表达的分析：根表达启动子的体外表达技术捕获和鉴定。

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Pseudomonas putida KT2440, a paradigm organism in biodegradation and a good competitive colonizer of the maize rhizosphere, was the subject of studies undertaken to establish the genetic determinants important for its rhizospheric lifestyle. By using in vivo expression technology (IVET) to positively select single cell survival, we identified 28 rap genes (root-activated promoters) preferentially expressed in the maize rhizosphere. The IVET system had two components: a mutant affected in aspartate-β-semialdehyde dehydrogenase (asd), which was unable to survive in the rhizosphere, and plasmid pOR1, which carries a promoterless asd gene. pOR1-borne transcriptional fusions of the rap promoters to the essential gene asd, which were integrated into the chromosome at the original position of the corresponding rap gene, were active and allowed growth of the asd strain in the rhizosphere. The fact that five of the rap genes identified in the course of this work had been formerly characterized as being related to root colonization reinforced the IVET approach. Up to nine rap genes encoded proteins either of unknown function or that had been assigned an unspecific role based on conservation of the protein family domains. Rhizosphere-induced fusions included genes with probable functions in the cell envelope, chemotaxis and motility, transport, secretion, DNA metabolism and defense mechanism, regulation, energy metabolism, stress, detoxification, and protein synthesis.
机译：恶臭假单胞菌 KT2440是生物降解的典范生物，是玉米根际的良好竞争定居者，是为确定对其根际生活重要的遗传决定因素而进行的研究课题。通过使用体内表达技术（IVET）积极选择单细胞存活率，我们鉴定了28个 rap 基因（ r oot- a 迷恋的 p romoters）优先在玉米根际表达。 IVET系统有两个组成部分：一个受天冬氨酸-β-半醛脱氢酶（ asd ）影响的突变体，该突变体无法在根际中存活;另一个质粒pOR1，其携带无启动子的 asd < / em>基因。 rap 启动子与必需基因 asd 的pOR1转录融合，整合到相应 rap 原始位置的染色体中基因，是活跃的并且允许 asd 菌株在根际中生长。在这项工作过程中鉴定出的五个 rap 基因以前被认为与根定植有关，这一事实加强了IVET方法。多达九个 rap 基因编码的蛋白质，要么功能未知，要么基于蛋白质家族结构域的保守性被赋予了非特异性作用。根际诱导的融合包括在细胞膜，趋化性和运动性，转运，分泌，DNA代谢和防御机制，调节，能量代谢，应激，排毒和蛋白质合成中可能具有功能的基因。

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