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Characterisation of the mgo operon in Pseudomonas syringae pv. syringae UMAF0158 that is required for mangotoxin production

机译:丁香假单胞菌pv中mgo操纵子的表征。生产芒果毒素所需的丁香香精UMAF0158

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摘要

In the present study, we performed a RT-PCR analysis, insertional inactivation mutagenesis, a promoter expression analysis and terminator localisation to study the gene cluster containing the mgoA gene. Additionally, we evaluated the importance of mgoC, mgoA and mgoD in mangotoxin production. A sequence analysis revealed an operon-like organisation. A promoter sequence was located upstream of the mgoB gene and was found to drive lacZ transcription. Two terminators were located downstream of the mgoD gene. RT-PCR experiments indicated that the four genes (mgoBCAD) constitute a transcriptional unit. This operon is similar in genetic organisation to those in the three other P. syringae pathovars for which complete genomes are available (P. syringae pv. syringae B728a, P. syringae pv. tomato DC3000 and P. syringae pv. phaseolicola 1448A). Interestingly, none of these three reference strains is capable of producing mangotoxin. Additionally, extract complementation resulted in a recovery of mangotoxin production when the defective mutant was complemented with wild-type extracts.
机译:在本研究中,我们进行了RT-PCR分析,插入失活诱变,启动子表达分析和终止子定位,以研究包含mgoA基因的基因簇。此外,我们评估了mgoC,mgoA和mgoD在芒果毒素生产中的重要性。序列分析显示出类似操纵子的组织。一个启动子序列位于mgoB基因的上游,并被发现可以驱动lacZ转录。两个终止子位于mgoD基因的下游。 RT-PCR实验表明,这四个基因(mgoBCAD)构成一个转录单位。该操纵子在遗传结构上与可用于其完整基因组的其他三个丁香假单胞菌致病病原体相似(丁香假单胞菌对丁香假单胞菌B728a,丁香假单胞菌对番茄DC3000和丁香假单胞菌对菜豆1448A)。有趣的是,这三个参考菌株均不能产生芒果毒素。另外,当缺陷突变体与野生型提取物互补时,提取物互补导致芒果毒素产生的恢复。

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