1999
DOI: 10.1016/s0168-6445(98)00033-3
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Strategies for isolation of in vivo expressed genes from bacteria

Abstract: The discovery and characterization of genes specifically induced in vivo upon infection and/or at a specific stage of the infection will be the next phase in studying bacterial virulence at the molecular level. Genes isolated are most likely to encode virulence-associated factors or products essential for survival, bacterial cell division and multiplication in situ. Identification of these genes is expected to provide new means to prevent infection, new targets for, antimicrobial therapy, as well as new insigh… Show more

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Cited by 15 publications
(18 citation statements)
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“…These methods have largely utilised recent advances in genomics and genomic based techniques. Many of these techniques aim to identify those genes encoding factors required for bacterial cell viability in vivo, which may not be essential for survival in vitro [20,25]. Several of these techniques are described below.…”
Section: Methods For Exploring Virulencementioning
confidence: 99%
“…These methods have largely utilised recent advances in genomics and genomic based techniques. Many of these techniques aim to identify those genes encoding factors required for bacterial cell viability in vivo, which may not be essential for survival in vitro [20,25]. Several of these techniques are described below.…”
Section: Methods For Exploring Virulencementioning
confidence: 99%
“…Therefore, an alternative approach to identify bacterial factors involved in infection processes could be based on the search for bacterial genes specifically up‐regulated in planta . Recently, a large number of bacterial genetic techniques have been developed leading to the identification and further analysis of factors essential for in vivo growth, such as IVET ( In Vivo Expression Technology) and RIVET (Reverse IVET) techniques (Slauch and Camilli, 2000), signature tagged mutagenesis (STM), and difference fluorescent induction (DFI), amongst others (Handfield and Levesque, 1999). Moreover, classical in vitro systems do not always represent an exact picture of what is happening in plant–bacteria interactions, because, in many cases, the infection processes are regulated and stimulated by host factors (Finlay and Falkow, 1997).…”
Section: Characterization Of Erwinia Chrysanthemi Host‐induced Clonesmentioning
confidence: 99%
“…1). The screening system was based on the use of reporter gene fusions to identify in vivo induced genes (Handfield and Levesque, 1999; Slauch and Camilli, 2000), and consisted of the following steps: (i) in vitro selection of non‐induced genes (white colonies in King's B (KB) plates); (ii) inoculation of these candidates in chicory discs and selection of in vivo induced ones after 24 h of incubation at 28 °C (blue chicory discs); and (iii) comprobation that the in vivo induced clones remained non‐inducible in vitro (white colonies in KB plates). Genomic DNA from the positive candidates was obtained using the Genomic prep Amersham kit, following the manufacturer's instructions (Amersham Pharmacia Biotech Inc., Piscataway, NJ, USA).…”
Section: Characterization Of Erwinia Chrysanthemi Host‐induced Clonesmentioning
confidence: 99%
“…It is generally accepted that genes of a pathogen that are specifically expressed during infection are likely to be important to pathogenicity. Numerous technologies such as in vivo expression technology (IVET), differential fluorescence induction (DFI) and signature‐tagged mutagenesis (STM) have been developed over the past decade to identify genes of pathogens that are specifically expressed during an actual infectious process (reviewed in [3]). Those methods have unfortunately not been useful for identifying in vivo induced genes of A. actinomycetemcomitans since no well‐validated and widely accepted animal model was available until very recently.…”
Section: Introductionmentioning
confidence: 99%