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利用体内表达技术对猪感染期间表达的沙门氏菌基因进行全基因组筛选。

Genome-wide screen of Salmonella genes expressed during infection in pigs, using in vivo expression technology.

作者信息

Huang Yanyan, Leming Christopher L, Suyemoto Mitsu, Altier Craig

机构信息

Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.

出版信息

Appl Environ Microbiol. 2007 Dec;73(23):7522-30. doi: 10.1128/AEM.01481-07. Epub 2007 Oct 5.

Abstract

Pigs are a food-producing species that readily carry Salmonella but, in the great majority of cases, do not show clinical signs of disease. Little is known about the functions required by Salmonella to be maintained in pigs. We have devised a recombinase-based promoter-trapping strategy to identify genes with elevated expression during pig infection with Salmonella enterica serovar Typhimurium. A total of 55 clones with in vivo-induced promoters were selected from a genomic library of approximately 10,000 random Salmonella DNA fragments fused to the recombinase cre, and the cloned DNA fragments were analyzed by sequencing. Thirty-one genes encoding proteins involved in bacterial adhesion and colonization (including bcfA, hscA, rffG, and yciR), virulence (metL), heat shock (hscA), and a sensor of a two-component regulator (hydH) were identified. Among the 55 clones, 19 were isolated from both the tonsils and the intestine, while 23 were identified only in the intestine and 13 only in tonsils. High temperature and increased osmolarity were identified as environmental signals that induced in vivo-expressed genes, suggesting possible signals for expression.

摘要

猪是一种产肉动物,很容易携带沙门氏菌,但在绝大多数情况下不会表现出疾病的临床症状。关于沙门氏菌在猪体内维持生存所需的功能,我们了解得很少。我们设计了一种基于重组酶的启动子捕获策略,以鉴定在猪感染肠炎沙门氏菌血清型鼠伤寒沙门氏菌期间表达上调的基因。从大约10,000个与重组酶cre融合的随机沙门氏菌DNA片段的基因组文库中,共筛选出55个具有体内诱导启动子的克隆,并对克隆的DNA片段进行测序分析。鉴定出31个编码参与细菌黏附和定植(包括bcfA、hscA、rffG和yciR)、毒力(metL)、热休克(hscA)以及双组分调节传感器(hydH)的蛋白质的基因。在这55个克隆中,19个是从扁桃体和肠道中都分离得到的,23个仅在肠道中鉴定到,13个仅在扁桃体中鉴定到。高温和渗透压升高被确定为诱导体内表达基因的环境信号,这表明它们可能是表达的信号。

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