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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Description of two new plasmids isolated from Francisella philomiragia strains and construction of shuttle vectors for the study of Francisella tularensis.
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Description of two new plasmids isolated from Francisella philomiragia strains and construction of shuttle vectors for the study of Francisella tularensis.

机译：从费氏弗朗西斯菌菌株中分离的两种新质粒的描述以及用于研究土拉弗朗西斯菌的穿梭载体的构建。

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摘要

Francisella tularensis is the causative agent of tularemia, a zoonotic disease often transmitted to humans by infected animals. The lack of useful specific genetic tools has long hampered the study of F. tularensis subspecies. We identified and characterized two new plasmids, pF242 and pF243, isolated from Francisella philomiragia strains ATCC 25016 and ATCC 25017, respectively. Sequence analysis revealed that pF242 and pF243 are closely related to pC194 and pFNL10 plasmids, respectively. Two generations of pF242- and pF243-based shuttle vectors, harboring several antibiotic resistance markers, were developed. We used the first generation to compare transformation efficiencies in two virulent F. tularensis subspecies. We found that electroporation was more efficient than cryotransformation: almost all vectors tested were successfully introduced by electroporation into Francisella strains with a high level of efficiency. The second generation of shuttle vectors, containing a multiple cloning site and/or gfp gene downstream of Francisella groES promotor, was used for GFP production in F. tularensis. The development of new shuttle vectors offers new perspectives in the genetic manipulation of F. tularensis, helping to elucidate the mechanisms underlying its virulence.
机译：图拉弗朗西斯菌是图拉血病的病原体，图拉血球病是一种人畜共患疾病，通常通过被感染的动物传播给人类。长期以来，缺乏有用的特异性遗传工具一直阻碍了杜鹃花亚种的研究。我们鉴定并鉴定了分别从费氏弗氏杆菌菌株ATCC 25016和ATCC 25017分离的两个新质粒pF242和pF243。序列分析表明pF242和pF243分别与pC194和pFNL10质粒密切相关。开发了两代基于pF242和pF243的穿梭载体，它们带有几种抗生素抗性标记。我们使用第一代技术来比较两个强毒的F. tularensis亚种的转化效率。我们发现电穿孔比冷冻转化更有效：几乎所有测试的载体都通过电穿孔成功地高效导入了弗朗西斯菌菌株。第二代穿梭载体包含弗朗西斯菌groES启动子下游的多个克隆位点和/或gfp基因，被用于图拉菌中的GFP生产。新的穿梭载体的开发为t.ensis的遗传操作提供了新的见解，有助于阐明其毒力的机制。

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