Single-step purification of recombinant Thermus aquaticus DNA polymerase using DNA-aptamer immobilized novel affinity magnetic beads

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dc.contributor.authorÖktem, Hüseyin Avni
dc.contributor.authorBayramoğlu, Gülay
dc.contributor.authorÖzalp, V. Cengiz
dc.contributor.authorArica, M. Yakup
dc.date.accessioned2020-06-25T17:44:02Z
dc.date.available2020-06-25T17:44:02Z
dc.date.issued2007
dc.descriptionOzalp, Veli Cengiz/0000-0002-7659-5990
dc.description.abstractA DNA aptamer specific for Thermus aquaticus DNA polymerase (Taq-polymerase) was immobilized on magnetic beads, which were prepared in the presented study. The effect of various parameters including pH, temperaturem and aptamer concentration on the immobilization of 5'-thiol labeled DNA-aptamer onto glutaric dialdhyde activated magnetic beads was evaluated. The binding conditions of Taq-polymerase on the aptamer immobilized magnetic beads were studied using commercial Taq-polymerase to characterize the surface complexation reaction. Efficiency of affinity magnetic beads in the purification of recombinant Taq-polymerase from crude extracts was also evaluated. For this case, the enzyme "recombinant Taq-DNA polymerase" was cloned and expressed using an Amersham E. coli GST-Gene Fusion Expression system. Crude extracts were in contact with affinity magnetic beads for 30 min and were collected by magnetic field application. The purity of the eluted Tag-polymerase from the affinity beads, as determined by HPLC, was 93% with a recovery of 89% in a one-step purification protocol. Apparently, the system was found highly effective as one step for the low-cost purification of Taq-polymerase in bacterial crude extract.en_US
dc.identifier.citationclosedAccessen_US
dc.identifier.doi10.1021/bp0602505
dc.identifier.endpage154en_US
dc.identifier.issn8756-7938
dc.identifier.issn1520-6033
dc.identifier.issue1en_US
dc.identifier.pmid17269682
dc.identifier.scopus2-s2.0-33847098767
dc.identifier.scopusqualityQ2
dc.identifier.startpage146en_US
dc.identifier.urihttps://doi.org10.1021/bp0602505
dc.identifier.urihttps://hdl.handle.net/20.500.12587/3989
dc.identifier.volume23en_US
dc.identifier.wosWOS:000243927600021
dc.identifier.wosqualityQ1
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.language.isoen
dc.publisherWileyen_US
dc.relation.ispartofBiotechnology Progress
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.titleSingle-step purification of recombinant Thermus aquaticus DNA polymerase using DNA-aptamer immobilized novel affinity magnetic beadsen_US
dc.typeArticle

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