A vaccinia virus isatin-beta-thiosemicarbazone resistance mutation maps in the viral gene encoding the 132-kDa subunit of RNA polymerase.
The mutation in a vaccinia virus mutant resistant to inhibition by isatin-beta-thiosemicarbazone was mapped by marker rescue. DNA from the resistant mutant was cloned into cosmid and plasmid vectors and tested for its ability to convert wild-type vaccinia virus to IBT resistant virus in a helper-mediated marker rescue protocol. Resistance was mapped in this way to a 0.9-kb DNA fragment derived from the HindIII A fragment of vaccinia genome. Southern blot hybridization using this DNA as a probe demonstrated that the 0.9-kb fragment is contained within the DNA sequence encoding the second largest subunit of vaccinia RNA polymerase, rpo132. Thus, mutation of rpo132 can cause resistance to IBT in vaccinia virus.
Condit
R C
RC
Department of Immunology and Medical Microbiology, University of Florida, Gainesville 32610-0266.
Easterly
R
R
Pacha
R F
RF
Fathi
Z
Z
Meis
R J
RJ
D015139:Blotting, Southern; D003001:Cloning, Molecular; D003360:Cosmids / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D004351:Drug Resistance / Q000235:genetics*; D005814:Genes, Viral / Q000235:genetics*; D007510:Isatin / Q000031:analogs & derivatives* / Q000494:pharmacology; D009154:Mutation / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D014616:Vaccinia virus / Q000187:drug effects / Q000235:genetics*
D015342:DNA Probes; C013578:isatin beta-thiosemicarbazone; D007510:Isatin; D012321:DNA-Directed RNA Polymerases