Comprehensive assignment of roles for Salmonella typhimurium genes in intestinal colonization of food-producing animals.
Comprehensive assignment of roles for Salmonella typhimurium genes in intestinal colonization of food-producing animals.
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DOI：
10.1371/journal.pgen.1003456
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发表时间：
2013-04
期刊：
影响因子：
4.5
通讯作者：
Stevens MP
中科院分区：
文献类型：
--
作者：
Chaudhuri RR;Morgan E;Peters SE;Pleasance SJ;Hudson DL;Davies HM;Wang J;van Diemen PM;Buckley AM;Bowen AJ;Pullinger GD;Turner DJ;Langridge GC;Turner AK;Parkhill J;Charles IG;Maskell DJ;Stevens MP
鸡、猪和牛是肠道沙门氏菌的主要宿主，肠道沙门氏菌是一种世界范围内重要的食源性病原体。虽然自第一个沙门氏菌基因组发表以来已经过去了十年，但仍有数千个基因具有假设或未知的功能，并且水库宿主的定植基础也不明确。此外，以前的调查的沙门氏菌基因在体内的作用集中在鼠伤寒模型的全身毒力，肠道持久性的遗传基础，从而人畜共患病传播的研究很少。因此，我们筛选了S.通过转座子定向插入位点测序（TraDIS）在鸡、猪和牛中检测肠道血清型鼠伤寒沙门氏菌。通过转座子侧翼区的大规模平行测序，同时分配每个宿主中7，702个突变体的身份和相对适合度。表型被分配给2，715个不同的基因，提供了前所未有的分辨率的表型-基因型图。这些数据是自洽的，因为观察到给定基因或途径中的多个独立突变产生类似的适应性成本。通过筛选鸡中确定的无效突变体进一步验证表型。我们的数据表明，一组核心的基因是必需的感染的所有三个宿主物种，和较小的基因组可能介导的持久性在特定的主机。通过对关键宿主中数千个沙门氏菌基因的作用进行分配，我们的数据有助于系统方法来了解发病机制和合理设计新型交叉保护疫苗和抑制剂。此外，通过同时分配在复杂池中筛选的超过90%的突变体的基因型和表型，我们的数据建立了TraDIS作为一个强大的工具，以最小的动物使用将丰富的功能注释应用于微生物基因组。 鼠伤寒沙门氏菌是人类肠道感染的主要原因，通常从鸡、猪、牛或其产品中获得。为了了解这些储存宿主中的持久性和发病机理的基础，并为新型疫苗和治疗的设计提供信息，我们生成了7，702个S的文库。鼠伤寒杆菌突变体，每个突变体在基因组中的随机位置上携带插入物。使用DNA测序，我们确定了每个突变体中被破坏的基因，并确定了其在实验室培养和小鼠，鸡，猪和牛的实验感染后的相对丰度。该方法允许同时研究大量突变体，大大减少了进行全面筛选所需的动物数量。我们发现了在培养中生长但在一种或多种动物中不能存活的突变体。在这些突变体中被破坏的基因被推断为对感染过程是重要的。这些基因中的大多数是所有三种食物生产动物所必需的，但较小的基因子集可能介导特定宿主物种中的持久性。这些数据提供了自然宿主中任何细菌病原体的毒力相关基因的最全面的图谱，并且与控制策略的设计高度相关。
Chickens, pigs, and cattle are key reservoirs of Salmonella enterica, a foodborne pathogen of worldwide importance. Though a decade has elapsed since publication of the first Salmonella genome, thousands of genes remain of hypothetical or unknown function, and the basis of colonization of reservoir hosts is ill-defined. Moreover, previous surveys of the role of Salmonella genes in vivo have focused on systemic virulence in murine typhoid models, and the genetic basis of intestinal persistence and thus zoonotic transmission have received little study. We therefore screened pools of random insertion mutants of S. enterica serovar Typhimurium in chickens, pigs, and cattle by transposon-directed insertion-site sequencing (TraDIS). The identity and relative fitness in each host of 7,702 mutants was simultaneously assigned by massively parallel sequencing of transposon-flanking regions. Phenotypes were assigned to 2,715 different genes, providing a phenotype–genotype map of unprecedented resolution. The data are self-consistent in that multiple independent mutations in a given gene or pathway were observed to exert a similar fitness cost. Phenotypes were further validated by screening defined null mutants in chickens. Our data indicate that a core set of genes is required for infection of all three host species, and smaller sets of genes may mediate persistence in specific hosts. By assigning roles to thousands of Salmonella genes in key reservoir hosts, our data facilitate systems approaches to understand pathogenesis and the rational design of novel cross-protective vaccines and inhibitors. Moreover, by simultaneously assigning the genotype and phenotype of over 90% of mutants screened in complex pools, our data establish TraDIS as a powerful tool to apply rich functional annotation to microbial genomes with minimal animal use. Salmonella Typhimurium is a major cause of human diarrhoeal infections, usually acquired from chickens, pigs, cattle, or their products. To understand the basis of persistence and pathogenesis in these reservoir hosts, and to inform the design of novel vaccines and treatments, we generated a library of 7,702 S. Typhimurium mutants, each bearing an insertion at a random position in the genome. Using DNA sequencing, we identified the disrupted gene in each mutant and determined its relative abundance in a laboratory culture and after experimental infection of mice, chickens, pigs, and cattle. The method allowed large numbers of mutants to be investigated simultaneously, drastically reducing the number of animals required to perform a comprehensive screen. We identified mutants that grow in culture but do not survive in one or more of the animals. The genes disrupted in these mutants are inferred to be important for the infection process. Most of these genes were required in all three food-producing animals, but smaller subsets of genes may mediate persistence in a specific host species. The data provide the most comprehensive map of virulence-associated genes for any bacterial pathogen in natural hosts and are highly relevant for the design of control strategies.
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影响因子：
3.1
作者：
Bispham, J;Tripathi, BN;Wallis, TS
通讯作者：
Wallis, TS
影响因子：
7
作者：
Langridge, Gemma C.;Phan, Minh-Duy;Turner, A. Keith
通讯作者：
Turner, A. Keith
影响因子：
6.7
作者：
Gal-Mor O;Gibson DL;Baluta D;Vallance BA;Finlay BB
通讯作者：
Finlay BB
DOI：
10.1073/pnas.120163297
发表时间：
2000-06-06
影响因子：
11.1
作者：
Datsenko, KA;Wanner, BL
通讯作者：
Wanner, BL
影响因子：
56.9
作者：
HENSEL, M;SHEA, JE;HOLDEN, DW
通讯作者：
HOLDEN, DW