1 Ocak 2013 Dergi makalesi Açık Erişim

Doruk, Tugrul; Gedik, Sedef Tunca

Dublin Core

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  <dc:creator>Doruk, Tugrul</dc:creator>
  <dc:creator>Gedik, Sedef Tunca</dc:creator>
  <dc:date>2013-01-01</dc:date>
  <dc: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.</dc:description>
  <dc:identifier>https://aperta.ulakbim.gov.trrecord/17289</dc:identifier>
  <dc:identifier>oai:zenodo.org:17289</dc:identifier>
  <dc:rights>info:eu-repo/semantics/openAccess</dc:rights>
  <dc:rights>http://www.opendefinition.org/licenses/cc-by</dc:rights>
  <dc:source>BIOLOGIA 68(3) 358-364</dc:source>
  <dc:title>An efficient gene deletion system for Bacillus thuringiensis</dc:title>
  <dc:type>info:eu-repo/semantics/article</dc:type>
  <dc:type>publication-article</dc:type>
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