1 Ocak 2013 Dergi makalesi Açık Erişim

Doruk, Tugrul; Gedik, Sedef Tunca

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{
  "@context": "https://schema.org/", 
  "@id": 17289, 
  "@type": "ScholarlyArticle", 
  "creator": [
    {
      "@type": "Person", 
      "affiliation": "Gebze Inst Technol, Dept Mol Biol & Genet, TR-41400 Gebze, Kocaeli, Turkey", 
      "name": "Doruk, Tugrul"
    }, 
    {
      "@type": "Person", 
      "affiliation": "Gebze Inst Technol, Dept Mol Biol & Genet, TR-41400 Gebze, Kocaeli, Turkey", 
      "name": "Gedik, Sedef Tunca"
    }
  ], 
  "datePublished": "2013-01-01", 
  "description": "It is not easy to manipulate biosynthetic genes of Bacillus thuringiensis since there is a powerful methyl-specific restriction system in this microorganism. In this study, a PCR-based system was used to delete polyphosphate kinase gene (ppk) of Bacillus thuringiensis israelensis (Bti) by replacing the wild-type gene with a cassette containing the apramycin resistance gene as selectable marker. lambda-Red was used to promote recombination in Escherichia coli between a PCR-amplified apramycin resistance cassette (linear deletion cassette selectable in E. coli and Bti) and Bti DNA on a plasmid. The isolated mutant plasmid was transferred to Bti by conjugation. Double cross-over transformants were screened for their antibiotic resistance and the mutation was proven by PCR, southern blot hybridization and RT-PCR. The described method, which uses the advantage of quick plasmid construction in E. coli and simple transformation of linear deletion cassette, is very useful to delete entire gene/genes of Bti without any polar effects on genes transcriptionally downstream.", 
  "headline": "An efficient gene deletion system for Bacillus thuringiensis", 
  "identifier": 17289, 
  "image": "https://aperta.ulakbim.gov.tr/static/img/logo/aperta_logo_with_icon.svg", 
  "license": "http://www.opendefinition.org/licenses/cc-by", 
  "name": "An efficient gene deletion system for Bacillus thuringiensis", 
  "url": "https://aperta.ulakbim.gov.tr/record/17289"
}