Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Vaccinia virus WR, complete genome
This strain is recommended by ATCC for use in the tests described in Fed. Spec. A-A-1438 where only the taxon is specified.
This tissue culture adapted WR strain of vaccinia is derived from ATCC VR-119 and has been shown to serve as an effective expression vector for foreign genes (Mackett et al., J. Virol., 1984)
Dr. Moss received the WR strain in 1966 from Dr. Salzman who obtained the virus as ATCC VR-119 from the ATCC. The tissue culture adapted strain WR has been widely distributed and used in constructing vectors for gene expression (J. Gen Virol, 1986).
Vaccination recommended for lab staff manipulating vaccinia, monkeypox, or cowpox viruses (CDC/NIH Biosafety in Microbiological and Biomedical Laboratories, 4th ed, 1999 - stock no. 017-040-00547-4).
Cell associated virus from infected HeLa cells
Temperature: 37°C
Recommendations for Infection: Plate cells 24 hours prior to infection and infect when cultures are 70-80% confluent. Remove medium and inoculate with a small volume of virus (e.g. 1 mL per 25 cm2) diluted in Virus growth medium: EMEM to provide an optimal MOI (e.g. 0.1). Adsorb 1-2 hours at 37°C in a humidified 5% CO2 atmosphere, rocking every 20-30 minutes to redistribute inoculum. End adsorption by adding virus growth medium.
Incubation: Incubate infected culture for 2-3 days at 37°C in a humidified 5% CO2 atmosphere, until CPE are well advance throughout.
Salzman NP, Sebring ED. Sequential formation of vaccinia virus proteins and viral deoxyribonucleic acid replication. J. Virol. 1: 16-23, 1967. PubMed: 4248263
Mackett M, et al. Vaccinia virus: a selectable eukaryotic cloning and expression vector. Proc. Natl. Acad. Sci. USA 79: 7415-7419, 1982. PubMed: 6296831
Nucleotide (GenBank) : AY243312.1 Vaccinia virus WR, complete genome
Nucleotide (GenBank) : S64006 E3L=double-stranded RNA-binding protein [vaccinia virus, WR, ATCC VR1354, Genomic, 633 nt].
