2012
DOI: 10.1128/aem.06865-11
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Genome Scanning for Conditionally Essential Genes in Salmonella enterica Serotype Typhimurium

Abstract: ABSTRACTAs more whole-genome sequences become available, there is an increasing demand for high-throughput methods that link genes to phenotypes, facilitating discovery of new gene functions. In this study, we describe a new version of the Tn-seq method involving a modified EZ:Tn5transposon for genome-wide and quantitative mapping of all insertions in a complex mutant library utilizing massively parallel Illumina sequencing. This Tn-seq method was app… Show more

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Cited by 68 publications
(59 citation statements)
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“…Should a similar pathway exist in Salmonella, the addition of taurocholate to the C-24 position would prevent such a reaction (19,52,53). Yet, if such genes exist in Salmonella and are necessary to process bile acids as a prerequisite for SPI-1 inactivation, they were not identified by our screen, suggesting that they are either essential (and perhaps conditionally essential for growth on bile-containing selective media) or functionally redundant (54,55). Similarly, our random and directed mutagenesis approaches failed to identify entry mechanisms for bile acids.…”
Section: Discussionmentioning
confidence: 65%
“…Should a similar pathway exist in Salmonella, the addition of taurocholate to the C-24 position would prevent such a reaction (19,52,53). Yet, if such genes exist in Salmonella and are necessary to process bile acids as a prerequisite for SPI-1 inactivation, they were not identified by our screen, suggesting that they are either essential (and perhaps conditionally essential for growth on bile-containing selective media) or functionally redundant (54,55). Similarly, our random and directed mutagenesis approaches failed to identify entry mechanisms for bile acids.…”
Section: Discussionmentioning
confidence: 65%
“…The Tn index of these genes is shown in Panels C and D of Figure 4. We further recorded the genes that Barquist and colleagues had originally selected (301 genes from S. Typhi and 299 for S. Typhimurium) which contained a reanalysis of the data from Langridge [9], as well as all genes previously identiied as 'essential' by Knuth [7], Khatiwari [10], Canals [11] and Thiele [13], regardless of whether such genes were successfully inactivated by others. This produced an 'all inclusive' list of 847 genes putatively essential for growth and survival, or under strong selection, in LB medium.…”
Section: Current Topics In Salmonella and Salmonellosismentioning
confidence: 99%
“…Some research groups selected for conditions more closely resembling natural conditions of infection, for instance growth at 42°C instead of 37°C, to resemble the body temperature of mice that S. Typhimurium would typically encounter, or in the presence of bile acid ( [10], work conducted with strain ATCC 14028). Exposure to low pH has also been tested [8].…”
Section: Essential Genes Based On Published Gene Inactivation Studiesmentioning
confidence: 99%
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“…For this, we used Tn-seq, a genome-wide screening method previously used for Streptococcus pneumoniae, Pseudomonas aeruginosa, Salmonella enterica serotype Typhimurium, and H. influenzae (7)(8)(9)(10)(11). We identified the H. influenzae carbonic anhydrase (HICA) gene to be absolutely essential for survival and growth of NTHi under ambient CO 2 conditions in vitro.…”
mentioning
confidence: 99%