Expression and self-assembly of cowpea chlorotic mottle virus-like particles in Pseudomonas fluorescens.
Coat protein of the cowpea chlorotic mottle virus (CCMV), a plant bromovirus, has been expressed in a soluble form in a prokaryote, Pseudomonas fluorescens, and assembled into virus-like particles (VLPs) in vivo that were structurally similar to the native CCMV particles derived from plants. The CCMV VLPs were purified by PEG precipitation followed by separation on a sucrose density gradient and analyzed by size exclusion chromatography, UV spectrometry, and transmission electron microscopy. DNA microarray experiments revealed that the VLPs encapsulated very large numbers of different host RNAs in a non-specific manner. The development of a P. fluorescens expression system now enables production of CCMV VLPs by bacterial fermentation for use in pharmaceutical or nanotechnology applications.
Phelps
Jamie P
JP
The Dow Chemical Company, Dowpharma, 5501 Oberlin Drive, San Diego, CA 92121, USA.
Dao
Philip
P
Jin
Hongfan
H
Rasochova
Lada
L
D017795:Bromovirus / Q000302:isolation & purification / Q000502:physiology*; D002499:Centrifugation, Density Gradient; D015967:Gene Expression Regulation, Viral; D020411:Oligonucleotide Array Sequence Analysis; D011551:Pseudomonas fluorescens / Q000821:virology*; D014771:Virion; D019065:Virus Assembly / Q000502:physiology*