Description

Type: Linear dsDNA enveloped poxvirus

Tropism: Vaccinia virus enters cells via macropinocytosis and unknown cellular receptors giving the virus a broad tropism (most mammalian cell types)

Gene Map of VV(Li)-RucGFP-hNIS-tRFP:

 

Proposed Uses

Standard titer stocks are ready to use in cell killing assays or for generation of new viral stocks. High-titer purified stocks are ready for immediate use in in vivo studies.

Protocol Tips

Propagation: Inoculate producer cells (e.g. Vero) at an MOI of 0.1 for 2-3 hours. After 48 hours harvest culture supernatant and cells and freeze-thaw or sonicate three times. Clarify the supernatant by low-speed centrifugation and purify through a  sucrose cushion.  Resuspend the viral pellet in 1mM TrisCl, pH 9.

Viral Titration: Titer virus by TCID₅₀/mL endpoint dilution assay. Score CPE positive cells based on the presence of dead cells 3-5 days after inoculation. Alternatively, titer viruses by plaque assay and crystal violet staining. Vero cells can be used for TCID₅₀ titrations.

Regulatory Information

Biosafety Level: 2

USDA transport permit required? Yes, you must have a USDA transport permit to ship Vaccinia virus. Please visit the USDA website to apply.