1 Ocak 2013 Dergi makalesi Açık Erişim

Doruk, Tugrul; Gedik, Sedef Tunca

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  <identifier identifierType="URL">https://aperta.ulakbim.gov.tr/record/17289</identifier>
  <creators>
    <creator>
      <creatorName>Doruk, Tugrul</creatorName>
      <givenName>Tugrul</givenName>
      <familyName>Doruk</familyName>
      <affiliation>Gebze Inst Technol, Dept Mol Biol &amp; Genet, TR-41400 Gebze, Kocaeli, Turkey</affiliation>
    </creator>
    <creator>
      <creatorName>Gedik, Sedef Tunca</creatorName>
      <givenName>Sedef Tunca</givenName>
      <familyName>Gedik</familyName>
      <affiliation>Gebze Inst Technol, Dept Mol Biol &amp; Genet, TR-41400 Gebze, Kocaeli, Turkey</affiliation>
    </creator>
  </creators>
  <titles>
    <title>An Efficient Gene Deletion System For Bacillus Thuringiensis</title>
  </titles>
  <publisher>Aperta</publisher>
  <publicationYear>2013</publicationYear>
  <dates>
    <date dateType="Issued">2013-01-01</date>
  </dates>
  <resourceType resourceTypeGeneral="Text">Journal article</resourceType>
  <alternateIdentifiers>
    <alternateIdentifier alternateIdentifierType="url">https://aperta.ulakbim.gov.tr/record/17289</alternateIdentifier>
  </alternateIdentifiers>
  <relatedIdentifiers>
    <relatedIdentifier relatedIdentifierType="DOI" relationType="IsIdenticalTo">10.2478/s11756-013-0184-4</relatedIdentifier>
  </relatedIdentifiers>
  <rightsList>
    <rights rightsURI="http://www.opendefinition.org/licenses/cc-by">Creative Commons Attribution</rights>
    <rights rightsURI="info:eu-repo/semantics/openAccess">Open Access</rights>
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  <descriptions>
    <description descriptionType="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.</description>
  </descriptions>
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