title: 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.
authors: Istituto per l'Ambiente Marino Costiero, UOS Capo Granitola-Consiglio Nazionale delle Ricerche, Trapani, Italy.
authors: Tiziana
authors: T
authors: Masullo
authors: Riccardo
authors: R
authors: Puccio
authors: Mariarosaria
authors: M
authors: Di Pierro
authors: Marcello
authors: M
authors: Tagliavia
authors: Paolo
authors: P
authors: Censi
authors: Valeria
authors: V
authors: Vetri
authors: Valeria
authors: V
authors: Militello
authors: Angela
authors: A
authors: Cuttitta
authors: Paolo
authors: P
authors: Colombo
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D003300:Copper / Q000032:analysis* / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D012615:Sea Anemones / Q000378:metabolism*
chemical_list: D049452:Green Fluorescent Proteins; D003300:Copper