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Development of a biosensor for copper detection in aqueous solutions using an Anemonia sulcata recombinant GFP.

Abstract：

:Fluorescent proteins from marine organisms represent potential candidates for biosensor development. In this paper, we described the isolation of a native green fluorescent protein from Anemonia sulcata and the cloning and purification of its equivalent as a recombinant protein in Escherichia coli. Furthermore, the spectroscopic behaviours of the native and recombinant GFPs were investigated as a function of Cu(2+), Cd(2+), Pb(2+) and Ni(2+) concentration. Our results suggest the high selectivity of both proteins at copper than the other metals and, for the recombinant protein, a great sensitivity at a very low concentration (0.1-1 μM). Moreover, starting from these data, using the combination of molecular biology techniques and optical setup, we developed a device for the detection of Cu(2+) in water solutions. The quenching effect detected with the device showed that the relative attenuation of the signal (0.46 ± 0.02 AU) was slightly larger than the data measured by fluorescence spectra (0.65 ± 0.03 AU). The good sensitivity in the span of two orders of the magnitude of Cu(2+) concentration, the fact that the instrument is made up of low-cost and sturdy parts and the selective quenching of rAsGFP to copper ions make this setup suited as a low cost, on-the-field, copper ion-specific biosensor.

journal_name

Appl Biochem Biotechnol

journal_title

Applied biochemistry and biotechnology

authors

Masullo T,Puccio R,Di Pierro M,Tagliavia M,Censi P,Vetri V,Militello V,Cuttitta A,Colombo P

doi

10.1007/s12010-013-0669-1

subject

Has Abstract

pub_date

2014-02-01 00:00:00

pages

2175-87

issue

4

eissn

0273-2289

issn

1559-0291

journal_volume

172

pub_type

杂志文章

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