Electrochemical detection of beta-1,3-glucanase gene from transgenic capsicums using asymmetric PCR generated by a detecting probe and an anchoring probe.
A design for recognition of beta-1,3-glucanase gene (Glu) specific sequence based on probe extension was described. The detecting probe DNA and the anchoring prober were hybridized with the same target DNA firstly, then the probes were extended by DNA polymerase reaction. After that the double strand DNA was denatured, and the extended detecting probe was immobilized on a glassy carbon electrode via nanoparticle gold (AuNP). In electrochemical detection (cyclic voltammetry, CV and differential pulse voltammetry, DPV), an increased peak current (i(p)) of the indicator (methylene blue, MB) was obtained compared with the probe without extension. Three differently long DNAs of Glu specific sequence were employed as the target: oligonucleotide acid, molecular cloning vector DNA and total genome DNA of transgenic capsicum. The estimated DPV detection limits for three targets of oligonucleotide, the molecular cloning vector DNA and genome DNA were 2.6x10(-13), 6.0x10(-13) and 8.0x10(-13)moll(-1) respectively.
Wu
Guofan
G
Northwest Normal University, Lanzhou, 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 / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D015342:DNA Probes; D018744:DNA, Plant / Q000737:chemistry / Q000302:isolation & purification*; D004566:Electrodes; D005822:Genetic Vectors; D043326:Glucan 1,3-beta-Glucosidase / Q000235:genetics*; D006046:Gold / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D015342:DNA Probes; D018744:DNA, Plant; D006046:Gold; D043326:Glucan 1,3-beta-Glucosidase