ArticleNucleic acids research2002
Identification of 113 conserved essential genes using a high-throughput gene disruption system in Streptococcus pneumoniae.
Article in Nucleic acids research, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 104 papers.
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Who cites it
104 citing papers in PubMed, 231 citations in OpenAlex.
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- The Bacterial Cell Wall: From Lipid II Flipping to Polymerization.Chemical reviews · 2022Review
- DEG 15, an update of the Database of Essential Genes that includes built-in analysis tools.Nucleic acids research · 2021Article
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- FtsZ-Ring Regulation and Cell Division Are Mediated by Essential EzrA and Accessory Proteins ZapA and ZapJ inFrontiers in microbiology · 2021Article
- The Pneumococcal Divisome: Dynamic Control ofFrontiers in microbiology · 2021Review
- tRNA methylation: An unexpected link to bacterial resistance and persistence to antibiotics and beyond.Wiley interdisciplinary reviews. RNA · 2020Review
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- Comprehensive dissection of dispensable genomic regions in Streptomyces based on comparative analysis approach.Microbial cell factories · 2020Article
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- Muraminomicins, novel ester derivatives: in vitro and in vivo antistaphylococcal activity.The Journal of antibiotics · 2019Article
44 more citing papers are in PubMed but not listed here.
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5 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
The recent availability of bacterial genome sequence information permits the identification of conserved genes that are potential targets for novel antibiotic drug discovery. Using a coupled bioinformatic/experimental approach, a list of candidate conserved genes was generated using a Microbial Concordance bioinformatics tool followed by a targeted disruption campaign. Pneumococcal sequence data allowed for the design of precise PCR primers to clone the desired gene target fragments into the pEVP3 'suicide vector'. An insertion-duplication approach was employed that used the pEVP3 constructs and resulted in the introduction of a selectable chloramphenicol resistance marker into the chromosome. In the case of non-essential genes, cells can survive the disruption and form chloramphenicol-resistant colonies. A total of 347 candidate reading frames were subjected to disruption analysis, with 113 presumed to be essential due to lack of recovery of antibiotic-resistant colonies. In addition to essentiality determination, the same high-throughput methodology was used to overexpress gene products and to examine possible polarity effects for all essential genes.
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