Comparative Genomics and Transduction Potential of Enterococcus faecalis Temperate Bacteriophages
Comparative Genomics and Transduction Potential of Enterococcus faecalis Temperate Bacteriophages
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DOI：
10.1128/jb.01293-09
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发表时间：
2010-02-01
影响因子：
3.2
通讯作者：
Horsburgh, Malcolm J.
中科院分区：
文献类型：
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作者：
Yasmin, Azra;Kenny, John G.;Horsburgh, Malcolm J.
为了确定温和的噬菌体的相对重要性，在水平基因转移的健身和毒力决定因素的粪肠球菌，一组47菌血症菌株进行了处理与诱导剂丝裂霉素C，诺氟沙星，紫外线辐射。从培养上清液中纯化了34个cDNA，并使用脉冲场凝胶电泳（PFGE）和限制性酶切图谱进行了区分。从这些分析中，八个代表性的基因组进行了焦磷酸测序，揭示了四个不同的组。发现三组基因组Phi FL 1至3具有序列相关性，其中Phi FL 1A至C以及Phi FL 2A和B具有最大的同一性（87至88%），而Phi FL 3A和B与Phi FL 1和2具有37至41%的同一性。Phi FL 4A与Phi FL 1至3具有3至12%的同一性。Phi FL 3A和B基因与乳酸乳球菌乳酸亚种的形态发生和裂解模块具有高度的DNA序列同一性。Cremoris原噬菌体。缓症链球菌血小板结合噬菌体尾蛋白PblA和PblB的同源物在每个测序的E.粪噬菌体很少有其他噬菌体基因编码潜在的毒力功能被确定，并有溶原性转化基因内溶素的远端，如已观察到的许多温和的基因组从机会病原体金黄色葡萄球菌和化脓性链球菌的载体的证据很少。E.使用八种细菌产生粪肠球菌JH 2 -2溶原菌，并检查这些溶原菌在大蜡螟中的相对适合度。几种溶原菌对G.与它们的同基因亲本相比。测试了八种质粒包装宿主DNA的能力，并且三种质粒显示出对实验室菌株OG 1 RF和JH 2 -2的幼稚宿主细胞的广泛转导非常有效。
To determine the relative importance of temperate bacteriophage in the horizontal gene transfer of fitness and virulence determinants of Enterococcus faecalis, a panel of 47 bacteremia isolates were treated with the inducing agents mitomycin C, norfloxacin, and UV radiation. Thirty-four phages were purified from culture supernatants and discriminated using pulsed-field gel electrophoresis (PFGE) and restriction mapping. From these analyses the genomes of eight representative phages were pyrosequenced, revealing four distinct groups of phages. Three groups of phages, Phi FL1 to 3, were found to be sequence related, with Phi FL1A to C and Phi FL2A and B sharing the greatest identity (87 to 88%), while Phi FL3A and B share 37 to 41% identity with Phi FL1 and 2. Phi FL4A shares 3 to 12% identity with the phages Phi FL1 to 3. The Phi FL3A and B phages possess a high DNA sequence identity with the morphogenesis and lysis modules of Lactococcus lactis subsp. cremoris prophages. Homologs of the Streptococcus mitis platelet binding phage tail proteins, PblA and PblB, are encoded on each sequenced E. faecalis phage. Few other phage genes encoding potential virulence functions were identified, and there was little evidence of carriage of lysogenic conversion genes distal to endolysin, as has been observed with genomes of many temperate phages from the opportunist pathogens Staphylococcus aureus and Streptococcus pyogenes. E. faecalis JH2-2 lysogens were generated using the eight phages, and these were examined for their relative fitness in Galleria mellonella. Several lysogens exhibited different effects upon survival of G. mellonella compared to their isogenic parent. The eight phages were tested for their ability to package host DNA, and three were shown to be very effective for generalized transduction of naive host cells of the laboratory strains OG1RF and JH2-2.