Development of a novel inducible bioluminescent and antibiotic resistance tagging system and its use to investigate the role of antibiotic production by Pectobacterium carotovorum ssp. carotovorum during potato tuber infection
Development of a novel inducible bioluminescent and antibiotic resistance tagging system and its use to investigate the role of antibiotic production by Pectobacterium carotovorum ssp. carotovorum during potato tuber infection
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开发一种新型诱导型生物发光和抗生素抗性标记系统，并将其用于研究胡萝卜果杆菌亚种抗生素生产的作用。
DOI：
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发表时间：
2009
影响因子：
1.8
通讯作者：
R. Fray
中科院分区：
文献类型：
--
作者：
K. Kovács;P. Hill;D. Grierson;C. Dodd;D. Pamfil;R. Fray
我们报告了一种新型Tn 7载体的构建，用于标记和计数来自复杂微生物群落的靶细菌。该系统利用可诱导生物发光和四环素抗性的盒，其整合在大多数革兰氏阴性菌种中存在的定义的中性位置。我们使用这种方法对果胶杆菌进行染色体标记，使得它可以在混合聚生体中进行计数，而不会对标记的菌株造成显著的生物负载。两个果胶杆菌菌株，碳青霉烯类抗生素生产商和同基因敲除菌株使用该系统进行标记。当感染的果胶杆菌是抗生素生产者（Car+）或碳青霉烯类抗生素生产者时，修饰的果胶杆菌菌株用于比较马铃薯块茎相关细菌和内生细菌利用果胶杆菌感染释放的营养物质在植物中获得优势和繁殖的程度。敲除（Car-）菌株。我们表明，在整个感染过程中，合成碳青霉烯的能力对果胶杆菌的数量有显着影响。虽然限制了其他细菌物种的数量，但碳青霉烯的生产允许果胶杆菌在腐烂的块茎中复制到更高的滴度。我们预计，Tn 7标记载体将用于其他研究人员在复杂的环境中研究生态相互作用。
We report the construction of a novel Tn7 vector for the tagging and enumeration of target bacteria from complex microbial communities. The system utilises a cassette for inducible bioluminescence and tetracycline resistance that integrates at a defined neutral position present in most Gram-negative species. We used this approach to chromosomally tag Pectobacterium such that it could be enumerated in mixed consortia without placing a significant bioburden on the tagged strain. Two Pectobacterium strains, a carbapenem antibiotic producer and an isogenic knock-out strain were tagged using this system. The modified Pectobacterium strains were used to compare the extent to which potato tuber-associated and endophytic bacteria can gain advantage and multiply in planta, utilising the nutrients released by a Pectobacterium infection, when the infecting Pectobacterium is either an antibiotic producer (Car+) or a carbapenem knock-out (Car−) strain. We show that the ability to synthesise carbapenem has a significant effect upon Pectobacterium numbers throughout the course of the infection. Whilst limiting the number of other bacterial species, carbepenem production allows the Pectobacterium to replicate to higher titres in the rotting tuber. We anticipate that the Tn7 tagging vector will be of use to other researchers studying ecological interactions in complex environments.
影响因子：
3.5
作者：
BERTRAND, KP;POSTLE, K;REZNIKOFF, WS
通讯作者：
REZNIKOFF, WS