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 journalArticle

14 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 1 2008

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Histidine
DNA
Proteins
Phosphatases
Alkaline Phosphatase
Recombinant proteins
Complexation
Recombinant Proteins
Limit of Detection
Complementary DNA

All Science Journal Classification (ASJC) codes

  • Biotechnology

Cite this

Conjugation of DNA with protein using His-tag chemistry and its application to the aptamer-based detection system. / Shimada, Josui; Maruyama, Tatsuo; Hosogi, Takuya; Tominaga, Jo; Kamiya, Noriho; Goto, Masahiro.

In: Biotechnology letters, Vol. 30, No. 11, 01.11.2008, p. 2001-2006.

Research output: Contribution to journalArticle

Shimada, Josui ; Maruyama, Tatsuo ; Hosogi, Takuya ; Tominaga, Jo ; Kamiya, Noriho ; Goto, Masahiro. / Conjugation of DNA with protein using His-tag chemistry and its application to the aptamer-based detection system. In: Biotechnology letters. 2008 ; Vol. 30, No. 11. pp. 2001-2006.
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