| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | ... CA, USA). 5. Insert specific primers for PCR diluted to 10 μM each, in either nuclease-free water or 1× TE (pH 8.0) (GenScript USA Inc., Piscataway, NJ, USA). 6. Dimethyl sulfoxide (DMSO) (Fisher Scientific, USA). 7. Luria Broth ... | Get A Quote |
In this chapter we demonstrate a method to produce virus-like particles (VLPs) from Escherichia coli. Standard bacterial protocols are used for the cloning, transformation, and expression of the protein subunits. A two-step protein purification method is highlighted: one step based on separating soluble proteins with ion-exchange affinity chromatography and a second polishing step using size-exclusion columns to isolate VLP species. The ensuing VLPs can be characterized with a variety of biophysical techniques including ultraviolet (UV)-visible spectroscopy for protein quantification, dynamic light scattering for size distribution determination, and transmission electron microscopy to ascertain size and morphol... More
