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  • Title: Conjugation of DNA with protein using His-tag chemistry and its application to the aptamer-based detection system.
    Author: Shimada J, Maruyama T, Hosogi T, Tominaga J, Kamiya N, Goto M.
    Journal: Biotechnol Lett; 2008 Nov; 30(11):2001-6. PubMed ID: 18604479.
    Abstract:
    We propose a novel method to prepare a DNA-protein conjugate using histidine-tag (His-tag) chemistry. Oligo-DNA was modified with nitrilotriacetate (NTA), which has high affinity to a His-tag on recombinant protein via the complexation of Ni(2+). Investigations using a microplate which displayed a complementary DNA-strand revealed that a NTA-modified DNA-protein conjugate was formed and immobilized in the presence of Ni(2+) on the microplate. We then adopted alkaline phosphatase (AP) as a model protein, and application of the DNA-AP conjugate was demonstrated in a thrombin aptamer-based detection system with a detection limit of approximately 10 nM.
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