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Doruk, Tugrul; Gedik, Sedef Tunca
<?xml version='1.0' encoding='UTF-8'?> <record xmlns="http://www.loc.gov/MARC21/slim"> <leader>00000nam##2200000uu#4500</leader> <datafield tag="700" ind1=" " ind2=" "> <subfield code="a">Gedik, Sedef Tunca</subfield> <subfield code="u">Gebze Inst Technol, Dept Mol Biol & Genet, TR-41400 Gebze, Kocaeli, Turkey</subfield> </datafield> <datafield tag="909" ind1="C" ind2="4"> <subfield code="p">BIOLOGIA</subfield> <subfield code="v">68</subfield> <subfield code="n">3</subfield> <subfield code="c">358-364</subfield> </datafield> <datafield tag="980" ind1=" " ind2=" "> <subfield code="a">user-tubitak-destekli-proje-yayinlari</subfield> </datafield> <datafield tag="540" ind1=" " ind2=" "> <subfield code="a">Creative Commons Attribution</subfield> <subfield code="u">http://www.opendefinition.org/licenses/cc-by</subfield> </datafield> <datafield tag="024" ind1=" " ind2=" "> <subfield code="a">10.2478/s11756-013-0184-4</subfield> <subfield code="2">doi</subfield> </datafield> <datafield tag="245" ind1=" " ind2=" "> <subfield code="a">An efficient gene deletion system for Bacillus thuringiensis</subfield> </datafield> <datafield tag="100" ind1=" " ind2=" "> <subfield code="a">Doruk, Tugrul</subfield> <subfield code="u">Gebze Inst Technol, Dept Mol Biol & Genet, TR-41400 Gebze, Kocaeli, Turkey</subfield> </datafield> <datafield tag="909" ind1="C" ind2="O"> <subfield code="o">oai:zenodo.org:17289</subfield> <subfield code="p">user-tubitak-destekli-proje-yayinlari</subfield> </datafield> <datafield tag="650" ind1="1" ind2="7"> <subfield code="2">opendefinition.org</subfield> <subfield code="a">cc-by</subfield> </datafield> <datafield tag="260" ind1=" " ind2=" "> <subfield code="c">2013-01-01</subfield> </datafield> <datafield tag="856" ind1="4" ind2=" "> <subfield code="u">https://aperta.ulakbim.gov.trrecord/17289/files/bib-6a1e474a-905f-4a15-b90c-090c41df1c06.txt</subfield> <subfield code="z">md5:73aa3b137fc4e24c0265599964730d74</subfield> <subfield code="s">117</subfield> </datafield> <datafield tag="542" ind1=" " ind2=" "> <subfield code="l">open</subfield> </datafield> <controlfield tag="005">20210315095243.0</controlfield> <controlfield tag="001">17289</controlfield> <datafield tag="980" ind1=" " ind2=" "> <subfield code="a">publication</subfield> <subfield code="b">article</subfield> </datafield> <datafield tag="520" ind1=" " ind2=" "> <subfield code="a">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.</subfield> </datafield> </record>
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