Intein-mediated site-specific conjugation of Quantum Dots to proteins in vivo

被引：21

作者：

Charalambous A. ^{[1
]}

Andreou M. ^{[1
]}

Skourides P.A. ^{[1
]}

机构：

[1] Department of Biological Sciences, University of Cyprus, 1678 Nicosia

来源：

Journal of Nanobiotechnology | / 7卷 / 1期

关键词：

Xenopus Embryo; Conjugation Method; Split Intein; Streptavidin Molecule; Superior Optical Property;

D O I：

10.1186/1477-3155-7-9

中图分类号：

学科分类号：

摘要：

We describe an intein based method to site-specifically conjugate Quantum Dots (QDs) to target proteins in vivo. This approach allows the covalent conjugation of any nanostructure and/or nanodevice to any protein and thus the targeting of such material to any intracellular compartment or signalling complex within the cells of the developing embryo. We genetically fused a pleckstrin-homology (PH) domain with the N-terminus half of a split intein (IN). The C-terminus half (I) of the intein was conjugated to QDs in vitro. IC-QD's and RNA encoding PH-IN were microinjected into Xenopus embryos. In vivo intein-splicing resulted in fully functional QD-PH conjugates that could be monitored in real time within live embryos. Use of Near Infra Red (NIR)-emitting QDs allowed monitoring of QD-conjugates within the embryo at depths where EGFP is undetectable demonstrating the advantages of QD's for this type of experiment. In conclusion, we have developed a novel in vivo methodology for the site-specific conjugation of QD's and other artificial structures to target proteins in different intracellular compartments and signaling complexes. © 2009 Charalambous et al; licensee BioMed Central Ltd.

引用

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