1 Ocak 2013 Dergi makalesi Açık Erişim

Doruk, Tugrul; Gedik, Sedef Tunca

Citation Style Language JSON

{
  "DOI": "10.2478/s11756-013-0184-4", 
  "abstract": "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.", 
  "author": [
    {
      "family": "Doruk", 
      "given": " Tugrul"
    }, 
    {
      "family": "Gedik", 
      "given": " Sedef Tunca"
    }
  ], 
  "container_title": "BIOLOGIA", 
  "id": "17289", 
  "issue": "3", 
  "issued": {
    "date-parts": [
      [
        2013, 
        1, 
        1
      ]
    ]
  }, 
  "page": "358-364", 
  "title": "An efficient gene deletion system for Bacillus thuringiensis", 
  "type": "article-journal", 
  "volume": "68"
}