Development of a biosensor for copper detection in aqueous solutions using an Anemonia sulcata recombinant GFP.
Appl Biochem Biotechnol
; 172(4): 2175-87, 2014 Feb.
Article
em En
| MEDLINE
| ID: mdl-24338300
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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Anêmonas-do-Mar
/
Técnicas Biossensoriais
/
Cobre
/
Proteínas de Fluorescência Verde
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
Idioma:
En
Revista:
Appl Biochem Biotechnol
Ano de publicação:
2014
Tipo de documento:
Article
País de afiliação:
Itália