Vaccinia Description and Custom Construct Information
Vaccinia virus is a member of the Orthopoxvirus genus. Recombinant vaccinia is a very useful vector because of its stability, ability to obtain high titer stocks ( up to 1010 pfu/ml), and its capacity to incorporate up to 40 kb of foreign DNA. Protein is expressed rapidly and at a high level. Vaccinia shows broad tropism in vivo and in vitro. 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. To aid in the recovery of the recombinant virus, plasmids have been generated that allow for selection of the recombinant vector. Since vaccinia encodes its own DNA-dependant RNA polymerase, which does not recognize foreign promoters, the cDNA is generally cloned downstream of a well defined virus promoter, close to an endogenous vaccinia promoter or use an endogenous 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.
Photo by Russell Kightley
Learn More About Vaccinia Products
All of our controls are available in 200ul aliquots
This virus will be provided in 1mM Tris, pH 9.0.
| Titer pfu/ml (plaque forming units/ml) | Volume |
| Ranges: 1E+08 pfu/ml to 1E+09 pfu/ml | 200ul |
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In-Stock Orders
You can browse/order our Vaccinia In-Stock Products here:
*Listed prices are for non-profit customers only, additional fees may apply.
**Please not, if you have used our ilabs system in the past, you will need to create a new account to order in-stock products from our new system
Custom Orders
***Custom vector orders still need to be submitted in iLabs
Most vectors with a catalog number are kept in stock but please call 319-335-6726 or email vectors@uiowa.edu for the latest availability as stock amounts are always changing.
Please allow 4-6 weeks for delivery of out-of-stock or "Upon Request" vectors.
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: $3,100 per construct plus shipping and handling.
Cost for MVA: $3,250 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: $3,700 per construct plus shipping and handling.
Cost for MVA: $3,900 per construct plus shipping and handling.