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Molecular Characterization and Lytic Activities of Streptococcus agalactiae Bacteriophages and Determination of Lysogenic-Strain Features

机译:无乳链球菌噬菌体的分子表征溶菌活性及溶源菌株特性的测定

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摘要

The application of mitomycin C induction to 114 genetically diverse Streptococcus agalactiae strains generated 36 phage suspensions. On electron microscopy of the phage suspensions, it was possible to assign the phages to the Siphoviridae family, with three different morphotypes (A, B, and C). Phage genetic diversity was evaluated by a PCR-based multilocus typing method targeting key modules located in the packaging, structural, host lysis, lysogeny, replication, and transcriptional regulation clusters and in the integrase genes and by DNA digestion with EcoRI, HindIII, and ClaI. Thirty-three phages clustering in six distantly related molecular phage groups (I to VI) were identified. Each molecular group was morphotype specific except for morphotype A phages, which were found in five of the six phage groups. The various phage groups defined on the basis of molecular group and morphotype had specific lytic activities, suggesting that each recognized particular host cell targets and had particular lytic mechanisms. Comparison of the characteristics of lysogenic and propagating strains showed no difference in the serotype or clonal complex (CC) identified by multilocus sequence typing. However, all the lysogenic CC17 and CC19 strains presented catabolic losses due to a lack of catabolic decay of dl-alpha-glycerol-phosphate substrates (CC17) and of alpha-d-glucose-1-phosphate (CC19). Moreover, the phages from CC17 lysogenic strains displayed lytic replication in bacterial hosts from all S. agalactiae phylogenetic lineages other than CC23, whereas phages obtained from non-CC17 lysogenic strains lysed bacteria of similar evolutionary origin. Our findings suggest that the adaptive evolution of S. agalactiae exposed the bacteria of this species to various phage-mediated horizontal gene transfers, which may have affected the fitness of the more virulent clones.
机译:将丝裂霉素C诱导应用于114个遗传多样性的无乳链球菌菌株中产生了36个噬菌体悬浮液。在噬菌体悬浮液的电子显微镜下,可以将噬菌体分配给具有三种不同形态型(A,B和C)的Siphoviridae家族。通过基于PCR的多基因座分型方法,针对位于包装,结构,宿主裂解,溶原性,复制和转录调控簇以及整合酶基因中的关键模块,并通过用EcoRI,HindIII和ClaI进行DNA消化,评估了噬菌体的遗传多样性。 。鉴定了在六个远距离相关的分子噬菌体组(I至VI)中的33个噬菌体。除了在六个噬菌体组中的五个发现的A型噬菌体外,每个分子组都是特定的形态型。基于分子基团和形态型定义的各种噬菌体基团具有特定的裂解活性,表明每个识别的噬菌体均具有特定的宿主细胞靶标并具有特定的裂解机制。溶原性和繁殖菌株的特征比较表明,通过多基因座序列分型鉴定的血清型或克隆复合体(CC)没有差异。然而,由于缺乏dl-α-甘油-磷酸底物(CC17)和α-d-葡萄糖-1-磷酸(CC19)的分解代谢衰减,所有溶原性CC17和CC19菌株均呈现分解代谢损失。而且,来自CC17溶原性菌株的噬菌体在除CC23以外的所有无乳链球菌系统发生谱系的细菌宿主中显示出裂解复制,而从非CC17溶原性菌株获得的噬菌体裂解了具有相似进化起源的细菌。我们的发现表明,无乳链球菌的适应性进化使该物种的细菌暴露于各种噬菌体介导的水平基因转移,这可能影响了更具毒性的克隆的适应性。

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