TY - JOUR
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.
UR - http://www.scopus.com/inward/record.url?scp=52449088063&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=52449088063&partnerID=8YFLogxK
U2 - 10.1007/s10529-008-9784-4
DO - 10.1007/s10529-008-9784-4
M3 - Article
C2 - 18604479
AN - SCOPUS:52449088063
SN - 0141-5492
VL - 30
SP - 2001
EP - 2006
JO - Biotechnology Letters
JF - Biotechnology Letters
IS - 11
ER -