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苏云金芽孢杆菌接合质粒pXO16介导的质粒捕获

Plasmid capture by the Bacillus thuringiensis conjugative plasmid pXO16.

作者信息

Timmery Sophie, Modrie Pauline, Minet Olivier, Mahillon Jacques

机构信息

Université Catholique de Louvain, Croix du Sud, Louvain-la-Neuve, Belgium.

出版信息

J Bacteriol. 2009 Apr;191(7):2197-205. doi: 10.1128/JB.01700-08. Epub 2009 Jan 30.

Abstract

Conjugation, mobilization, and retromobilization are three related mechanisms of horizontal gene transfer in bacteria. They have been extensively studied in gram-negative species, where retromobilization, the capture of DNA from a recipient by a donor cell, was shown to result from two successive steps: the transfer of the conjugative plasmid from the donor to the recipient followed by the retrotransfer of the mobilizable plasmid to the donor. This successive model was established for gram-negative bacteria but was lacking experimental data from the gram-positive counterparts. In the present work, the mobilization and retromobilization abilities of the conjugative plasmid pXO16 from Bacillus thuringiensis subsp. israelensis were studied using the mobilizable plasmids pUB110 and pE194 and the "nonmobilizable" element pC194 lacking the mob and oriT features (all from Staphylococcus aureus). Experimental data suggested a successive model, since different retromobilization frequencies were observed between the small plasmids. More importantly, retromobilization was shown to be delayed by 50 and 150 min for pUB110 and pE194, respectively, compared to pXO16 conjugation. Natural liquid foods (cow milk, soy milk, and rice milk) were used to evaluate the putative ecological impact of these transfers. In cow and soy milk, conjugation, mobilization, and retromobilization were shown to occur at frequencies of 8.0 x 10(-1), 1.0 x 10(-2), and 1.2 x 10(-4) transconjugants per recipient, respectively. These data are comparable to those obtained with LB medium and about 10-fold lower than in the case of rice milk. Taken together, these results emphasize the potential role of plasmid capture played by B. thuringiensis in natural environments.

摘要

接合、迁移和反向迁移是细菌中水平基因转移的三种相关机制。它们在革兰氏阴性菌中得到了广泛研究,在革兰氏阴性菌中,反向迁移(供体细胞从受体捕获DNA)被证明是由两个连续步骤导致的:接合性质粒从供体转移到受体,随后可移动质粒反向转移到供体。这个连续模型是针对革兰氏阴性菌建立的,但缺乏革兰氏阳性菌对应情况的实验数据。在本研究中,利用可移动质粒pUB110和pE194以及缺乏mob和oriT特征的“不可移动”元件pC194(均来自金黄色葡萄球菌),研究了苏云金芽孢杆菌以色列亚种的接合性质粒pXO16的迁移和反向迁移能力。实验数据表明存在一个连续模型,因为在小质粒之间观察到了不同的反向迁移频率。更重要的是,与pXO16接合相比,pUB110和pE194的反向迁移分别延迟了50分钟和150分钟。使用天然液体食品(牛奶、豆浆和米糊)来评估这些转移可能产生的生态影响。在牛奶和豆浆中,接合、迁移和反向迁移的发生频率分别为每受体8.0×10⁻¹、1.0×10⁻²和1.2×10⁻⁴个接合子。这些数据与在LB培养基中获得的数据相当,比在米糊中的情况低约10倍。综上所述,这些结果强调了苏云金芽孢杆菌在自然环境中质粒捕获的潜在作用。

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