A streamlined method for transposon mutagenesis of Rickettsia parkeri yields numerous mutations that impact infection.
A streamlined method for transposon mutagenesis of Rickettsia parkeri yields numerous mutations that impact infection.
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DOI：
10.1371/journal.pone.0197012
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Welch MD
中科院分区：
文献类型：
--
作者：
Lamason RL;Kafai NM;Welch MD
立克次体是专性胞内α-变形菌，在节肢动物和哺乳动物宿主中都表现出复杂的感染性生活史。立克次体作为专性胞内细菌，在哺乳动物感染期间高度适应于生活在各种宿主细胞内，包括血管内皮细胞。虽然人们认为立克次体产生了许多毒力因子，这些毒力因子可以侵占或破坏各种宿主细胞途径，但通过基因操作来识别导致感染的关键细菌因子一直是一个挑战。为了克服这一挑战，我们试图扩大现有的立克次体功能丧失突变体的库，使用改进的水手为基础的转座子诱变方案。在这里，我们提出了超过100个转座子突变体的斑点热组物种立克次体的分离。转座子插入破坏了其产物涉及多种途径的基因，包括细菌复制和代谢、IV型分泌系统、在宿主细胞相互作用和发病机制中具有先前确定的作用的因子，或具有未知功能的因子。鉴于需要确定关键的毒力因子，正向遗传筛选，如这将提供一个很好的平台，更直接地调查立克次体的生物学和发病机制。
The rickettsiae are obligate intracellular alphaproteobacteria that exhibit a complex infectious life cycle in both arthropod and mammalian hosts. As obligate intracellular bacteria, rickettsiae are highly adapted to living inside a variety of host cells, including vascular endothelial cells during mammalian infection. Although it is assumed that the rickettsiae produce numerous virulence factors that usurp or disrupt various host cell pathways, they have been challenging to genetically manipulate to identify the key bacterial factors that contribute to infection. Motivated to overcome this challenge, we sought to expand the repertoire of available rickettsial loss-of-function mutants, using an improved mariner-based transposon mutagenesis scheme. Here, we present the isolation of over 100 transposon mutants in the spotted fever group species Rickettsia parkeri. Transposon insertions disrupted genes whose products are implicated in a variety of pathways, including bacterial replication and metabolism, the type IV secretion system, factors with previously established roles in host cell interactions and pathogenesis, or are of unknown function. Given the need to identify critical virulence factors, forward genetic screens such as this will provide an excellent platform to more directly investigate rickettsial biology and pathogenesis.
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影响因子：
15.9
作者：
Al-Khodor S;Price CT;Kalia A;Abu Kwaik Y
通讯作者：
Abu Kwaik Y
影响因子：
3.7
作者：
Gillespie, Joseph J.;Williams, Kelly;Shukla, Maulik;Snyder, Eric E.;Nordberg, Eric K.;Ceraul, Shane M.;Dharmanolla, Chitti;Rainey, Daphne;Soneja, Jeetendra;Shallom, Joshua M.;Vishnubhat, Nataraj Dongre;Wattam, Rebecca;Purkayastha, Anjan;Czar, Michael;Crasta, Oswald;Setubal, Joao C.;Azad, Abdu F.;Sobral, Bruno S.
通讯作者：
Sobral, Bruno S.
影响因子：
3.7
作者：
Gillespie JJ;Ammerman NC;Dreher-Lesnick SM;Rahman MS;Worley MJ;Setubal JC;Sobral BS;Azad AF
通讯作者：
Azad AF
影响因子：
3.3
作者：
Gillespie, Joseph J.;Phan, Isabelle Q. H.;Azad, Abdu F.
通讯作者：
Azad, Abdu F.
影响因子：
3.2
作者：
Kaur, Simran J.;Rahman, M. Sayeedur;Azad, Abdu F.
通讯作者：
Azad, Abdu F.