Conjugation of fluorescent proteins with DNA oligonucleotides.
This work describes the synthesis of covalent ssDNA conjugates of six fluorescent proteins, ECFP, EGFP, E(2)GFP, mDsRed, Dronpa, and mCherry, which were cloned with an accessible C-terminal cystein residue to enable site-selective coupling using a heterobispecific cross-linker. The resulting conjugates revealed similar fluorescence emission intensity to the unconjugated proteins, and the functionality of the tethered oligonucleotide was proven by specific Watson-Crick base pairing to cDNA-modified gold nanoparticles. Fluorescence spectroscopy analysis indicated that the fluorescence of the FP is quenched by the gold particle, and the extent of quenching varied with the intrinsic spectroscopic properties of FP as well as with the configuration of surface attachment. Since this study demonstrates that biological fluorophores can be selectively incorporated into and optically coupled with nanoparticle-based devices, applications in DNA-based nanofabrication can be foreseen.
Lapiene
Vidmantas
V
Technische Universitat Dortmund, Fakultat Chemie, Biologisch-Chemische Mikrostrukturtechnik, Otto-Hahn Str. 6, D-44227 Dortmund, Germany.
Kukolka
Florian
F
Kiko
Kathrin
K
Arndt
Andreas
A
Niemeyer
Christof M
CM
D003001:Cloning, Molecular; D003432:Cross-Linking Reagents; D004277:DNA, Single-Stranded / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D005818:Genetic Engineering; D008164:Luminescent Proteins / Q000737:chemistry* / Q000235:genetics*; D009154:Mutation; D009841:Oligonucleotides / Q000737:chemistry*
D003432:Cross-Linking Reagents; D004277:DNA, Single-Stranded; D008164:Luminescent Proteins; D009841:Oligonucleotides