6533b7defe1ef96bd12768e7
RESEARCH PRODUCT
Properties of baculovirus particles displaying GFP analyzed by fluorescence correlation spectroscopy.
Jouni ToivolaKirsi OjalaPatrik O. MichelMatti VuentoChristian Oker-blomsubject
PhotochemistryvirusesClinical BiochemistryDetergentsGreen Fluorescent ProteinsFluorescence correlation spectroscopySpodopteraBiochemistryGreen fluorescent proteinDiffusionchemistry.chemical_compoundViral envelopeAnimalsSodium dodecyl sulfateMolecular BiologybiologyChemistryViral membranebiology.organism_classificationFluorescenceFusion proteinMolecular biologyMolecular WeightAutographa californicaLuminescent ProteinsSpectrometry FluorescenceElectrophoresis Polyacrylamide GelIndicators and ReagentsBaculoviridaeViral Fusion ProteinsAlgorithmsdescription
Abstract Recombinant baculovirus particles displaying green fluorescent protein (GFP) fused to the major envelope glycoprotein gp64 of the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) were characterized by fluorescence correlation spectroscopy (FCS). FCS detected Brownian motion of single, intact recombinant baculovirus display particles with a diffusion coefficient (D) of (2.89±0.74)10 8 cm2s 1 and an apparent hydrodynamic radius of 83.35±21.22 nm. In the presence of sodium dodecyl sulfate (SDS), Triton X-100, and octylglucoside, the diffusion time was reduced to the 0.2 ms range (D = 7.5710 7 cm2s 1), showing that the fusion proteins were anchored in the viral envelope. This allowed for a calculation of the number of single gp64 fusion proteins incorporated in the viral membrane. A mean value of 3.2 fluorescent proteins per virus particle was obtained. Our results show that FCS is the method of choice for studying enveloped viruses such as a display virus with one component being GFP.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2003-01-30 | Biological chemistry |