An electrochemical assay of beta-1,3-glucanase gene from transgenic capsicum using asymmetric PCR.
5'-Thiol-derivatized specific DNA probes were added to the single primer polymerase chain reaction (asymmetric PCR) solution. In the PCR process, the DNA probes extended in the presence of target; the extended probes were then immobilized on a glassy carbon electrode (GCE) via gold nanoparticles. Finally, methylene blue and the extended probes were combined and the electrochemical signals were measured. This signal was higher than that of the GCE modified only by the original probe. When there was no target in PCR solution, the probe did not extend and the signal did not increase. The specific sequences of the beta-1,3-glucanase gene were detected successfully from three targets with different length: oligonucleotide, molecule clone vector DNA, and total genome DNA of transgenic capsicum. The detection limits of 2.6 x 10(-13), 7.8 x 10(-13), and 9.1 x 10(-13) moll(-1) for oligonucleotide, molecule clone vector DNA, and total transgenic capsicum genome DNA were estimated.
Wu
Guofan
G
College of Chemistry and Chemical Engineering, Northwest Normal University, Lanzhou, Gansu Province, PR China. wugf1971@163.com
Wang
Zhihua
Z
Zhang
Huini
H
Yang
Ning
N
Du
Jie
J
Lu
Xiaoquan
X
Kang
Jingwan
J
D002212:Capsicum / Q000235:genetics*; D002623:Chemistry Techniques, Analytical; D004563:Electrochemistry; D043326:Glucan 1,3-beta-Glucosidase / Q000737:chemistry / Q000235:genetics*; D057230:Limit of Detection; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction
D043326:Glucan 1,3-beta-Glucosidase