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Custom Vaccinia Virus Constructs

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Please view our ordering information for custom constructs to learn how to register for an account, place orders for in-stock vectors and make payment through our online system.

Description

The recombinant vaccinia virus is produced by homologous recombination between a plasmid and the viral DNA within infected cells. The gene of interest is inserted in a non-essential locus of the vaccinia DNA or between two essential genes to minimize unstable recombinants. To aid in the recovery of the recombinant virus, plasmids have been generated that allow for selection of the recombinant virus.

Since vaccinia encodes its own DNA-dependant RNA polymerase, which does not recognize foreign promoters, the cDNA must be cloned downstream of a well defined virus promoter or close to an endogenous vaccinia promoter.

Vaccinia viruses are widely used for the study of poxvirus molecular biology, functional characterization and over expression of proteins, and as candidates for live vaccines or vaccine research.

Please check the safety considerations required by your institution. Many institutions require small pox vaccination prior to use for wild type vaccinia but not for MVA (Modified Vaccinia Ankara).

Amplification, purification and titer from viral particles

Material provided: 8ml of purified virus at a concentration ranging from 5 x107 pfu/ml to 1 x109 pfu/ml.

Sample required: Viral particles

Timeline: 3-4 weeks from the time the virus is received.

Services include:

  • Large-scale amplification
  • Purification by sucrose cushion
  • With every new lot an infectious titer (pfu/ml) is provided

Vehicle: Vaccinia vectors are re-suspended in 1mM Tris HCI pH 9.0.

Cost for wild type: $2,860 per construct plus shipping and handling.

Cost for MVA: $3,080 per construct plus shipping and handling.

Construction, selection, purification, amplification and titer

Material provided: 8ml of purified virus at a concentration ranging from 5 x107 pfu/ml to 1 x109 pfu/ml.

Sample required: 20 µg of vaccinia shuttle plasmid expressing the gene of interest at a concentration greater than 0.25 µg/µl.

Timeline: 6-8 weeks from the time the plasmid is received.

Services include:

  • Transfection
  • Cell sorting using a fluorescent marker for faster isolation of recombinants (MVA only)
  • Three or more rounds of selection depending on choice of shuttle plasmid
  • Large-scale amplification
  • Purification by sucrose cushion
  • With every new lot an infectious titer (pfu/ml) is provided

Vehicle: Vaccinia vectors are re-suspended in 1mM Tris HCI pH 9.0.

Cost for Wild type: $3,500 per construct plus shipping and handling.

Cost for MVA: $3,700 per construct plus shipping and handling.