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

doi:10.1007/s10529-008-9784-4

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

Applied Chemistry

Research output: Contribution to journal › Article › peer-review

24 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 language	English
Pages (from-to)	2001-2006
Number of pages	6
Journal	Biotechnology letters
Volume	30
Issue number	11
DOIs	https://doi.org/10.1007/s10529-008-9784-4
Publication status	Published - Nov 2008

All Science Journal Classification (ASJC) codes

Biotechnology
Bioengineering
Applied Microbiology and Biotechnology

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10.1007/s10529-008-9784-4

Cite this

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

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.",

author = "Josui Shimada and Tatsuo Maruyama and Takuya Hosogi and Jo Tominaga and Noriho Kamiya and Masahiro Goto",

note = "Funding Information: Acknowledgements The present work is supported by a Grant-in-Aid for the Global COE Program, {\textquoteleft}{\textquoteleft}Science for Future Molecular Systems{\textquoteright}{\textquoteright} from the Ministry of Education, Culture, Sports, Science and Technology of Japan.",

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T1 - Conjugation of DNA with protein using His-tag chemistry and its application to the aptamer-based detection system

AU - Shimada, Josui

AU - Maruyama, Tatsuo

AU - Hosogi, Takuya

AU - Tominaga, Jo

AU - Kamiya, Noriho

AU - Goto, Masahiro

N1 - Funding Information: Acknowledgements The present work is supported by a Grant-in-Aid for the Global COE Program, ‘‘Science for Future Molecular Systems’’ from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

PY - 2008/11

Y1 - 2008/11

N2 - 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.

AB - 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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EP - 2006

JO - Biotechnology Letters

JF - Biotechnology Letters

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

Abstract

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