Conjugation of DNA with protein using His-tag chemistry and its application to the aptamer-based detection system

Josui Shimada, Tatsuo Maruyama, Takuya Hosogi, Jo Tominaga, Noriho Kamiya, Masahiro Goto

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

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 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.

Original languageEnglish
Pages (from-to)2001-2006
Number of pages6
JournalBiotechnology letters
Volume30
Issue number11
DOIs
Publication statusPublished - Nov 2008

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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