Conjugation of DNA with protein using His-tag chemistry and its application to the aptamer-based detection system |
| |
| Authors: | Josui Shimada Tatsuo Maruyama Takuya Hosogi Jo Tominaga Noriho Kamiya Masahiro Goto |
| |
| Affiliation: | (1) Department of Applied Chemistry, Graduate School of Engineering, Center for Future Chemistry, Kyushu University, 744 Moto-oka, Fukuoka 819-0395, Japan;(2) Department of Chemical Science and Engineering, Kobe University, 1-1 Rokkodai, Kobe 657-8501, Japan |
| |
| 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. |
| |
| Keywords: | DNA aptamer DNA– protein conjugate Enzymatic reaction Histidine-tag Protein immobilization |
| 本文献已被 PubMed SpringerLink 等数据库收录！ |
|
