Work

Assembly of Hybrid Bacteriophage Qβ Virus-like Particles

Year: 2009

Type: article

Abstract: Bacteriophage Qbeta coat protein forms uniform virus-like particles when expressed recombinantly in a variety of organisms. We have inserted the IgG-binding Z domain at the carboxy terminus of the coat protein and coexpressed this chimeric subunit with native coat protein to create hybrid, IgG-binding virus-like particles. Extracellular osmolytes were found to have an effect on the efficiency of incorporation of fusion proteins into VLPs in Escherichia coli when a carbenicillin, but not a kanamycin, selection marker was used. The addition of sucrose to the growth medium decreased the incorporation efficiency; the osmoprotectant glycine betaine eliminated this effect. The decrease in efficiency was not observed when carbenicillin was omitted from the final expression culture. The addition of sodium chloride instead of sucrose gave rise to particles with a larger number of fusion proteins than the standard conditions. These results illustrate that cellular conditions should be taken into account even in apparently simple systems when natural or engineered protein nanoparticles are made.

Source: Biochemistry

DOI: 10.1021/bi901306p

Authors: Steven D. Brown, Jason D. Fiedler, M. G. Finn

Institution: Scripps Research Institute

Language: English

Topic: Bacteriophages and microbial interactions

Subfield: Ecology

Field: Environmental Science

Domain: Physical Sciences

Open Access status: green

Funders: National Institute of General Medical Sciences, National Center for Research Resources

Awards: Electron Dense Labels for Macromolecular Imaging, Resistance Driven Structural Design for HIV Therapies, G4796118608, G7086953010