The gene encoding the late nonstructural 36K protein of vaccinia virus is essential for virus reproduction.
Two genetic markers--the thymidine kinase gene of herpes simplex virus, and the beta-galactosidase gene of Escherichia coli--were incorporated into the 36K protein gene (IL1 gene according to the nomenclature of the Copenhagen strain of vaccinia virus; Goebel et al., 1990) from the HindIII-P DNA fragment of the LIVP strain (variant of Lister strain) of vaccinia virus (VV). After recombination of the obtained integration plasmid pVZ64-TK with the VV genome (tk-), it was found that the resultant TK+ viruses were unstable with respect to the Lac+ phenotype. On the basis of hybridization of DNA fragments of selected clones, a scheme for the formation of hybrid viruses is proposed, and an approach to a simple phenotypical discrimination between essential and non-essential genes for VV viability is described.
Shchelkunov
S N
SN
Institute of Molecular Biology, NPO Vector, Koltsovo, Novosibirsk region, Russia.
Ryazankina
O I
OI
Gashnikov
P V
PV
D000818:Animals; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004279:DNA, Viral; D016679:Genome, Viral; D010641:Phenotype; D010957:Plasmids; D051381:Rats; D011995:Recombination, Genetic; D014616:Vaccinia virus / Q000235:genetics / Q000502:physiology*; D017361:Viral Nonstructural Proteins / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics*; D016662:Virus Integration; D014779:Virus Replication / Q000235:genetics*
D004279:DNA, Viral; C066517:Vaccinia virus late protein K36; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins