1 Ocak 2013 Dergi makalesi Açık Erişim

Doruk, Tugrul; Gedik, Sedef Tunca

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    "creators": [
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        "affiliation": "Gebze Inst Technol, Dept Mol Biol & Genet, TR-41400 Gebze, Kocaeli, Turkey", 
        "name": "Doruk, Tugrul"
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      {
        "affiliation": "Gebze Inst Technol, Dept Mol Biol & Genet, TR-41400 Gebze, Kocaeli, Turkey", 
        "name": "Gedik, Sedef Tunca"
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    "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.", 
    "doi": "10.2478/s11756-013-0184-4", 
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    "journal": {
      "issue": "3", 
      "pages": "358-364", 
      "title": "BIOLOGIA", 
      "volume": "68"
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    "publication_date": "2013-01-01", 
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    "title": "An efficient gene deletion system for Bacillus thuringiensis"
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