Sensitively electrochemical sensing for sequence-specific detection of phosphinothricin acetyltransferase gene: Layer-by-layer films of poly-l-lysine and Au-carbon nanotube hybrid

Abstract

An electrochemical DNA sensing film was constructed based on the multilayers comprising of poly-l-lysine (pLys) and Au-carbon nanotube (Au-CNT) hybrid. A precursor film of mercaptopropionic acid (MPA) was firstly self-assembled on the Au electrode surface. pLys and Au-CNT hybrid layer-by-layer assembly films were fabricated by alternately immersing the MPA-modified electrode into the pLys solution and Au-CNT hybrid solution. Cyclic voltammetry was used to monitor the consecutive growth of the multilayer films by utilizing [Fe(CN)6]3-/4- and [Co(phen)3]3+/2+ as the redox indicators. The outer layer of the multilayer film was the positively charged pLys, on which the DNA probe was easily linked due to the strong electrostatic affinity. The hybridization detection of DNA was accomplished by using methylene blue (MB) as the indicator, which possesses different affinities to dsDNA and ssDNA. Differential pulse voltammetry was employed to record the signal response of MB and determine the amount of the target DNA sequence. The established biosensor has high sensitivity, a relatively wide linear range from 1.0 × 10-10 mol/L to 1.0 × 10-6 mol/L and the ability to discriminate the fully complementary target DNA from single or double base-mismatched DNA. The sequence-specific DNA related to phosphinothricin acetyltransferase gene from the transgenically modified plants was successfully detected. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.