Öktem, Hüseyin AvniBayramoğlu, GülayÖzalp, V. CengizArica, M. Yakup2020-06-252020-06-252007closedAccess8756-79381520-6033https://doi.org10.1021/bp0602505https://hdl.handle.net/20.500.12587/3989Ozalp, Veli Cengiz/0000-0002-7659-5990A 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.eninfo:eu-repo/semantics/closedAccessArticle23114615410.1021/bp06025052-s2.0-3384709876717269682Q2WOS:000243927600021Q1