Journal
BIOTECHNOLOGY LETTERS
Volume 30, Issue 11, Pages 2001-2006Publisher
SPRINGER
DOI: 10.1007/s10529-008-9784-4
Keywords
DNA aptamer; DNA-protein conjugate; enzymatic reaction; histidine-tag; protein immobilization
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Funding
- Ministry of Education, Culture, Sports, Science and Technology of Japan
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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 Ni2+. 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 Ni2+ 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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