Inducible transformation of fibroblasts using a metallothionein-v-myc gene construct.
An inducible oncogene construct has been engineered by coupling the MC29 v-myc oncogene to the sheep metallothionein promoter. Transfection of this plasmid, which also contains the neomycin resistance gene, into Rat-1 cells, has resulted in the isolation of independent clones resistant to G418. Certain of these clones were found to exhibit inducible transformation in response to ZnSO4. Transformation was graded with increasing ZnSO4 levels and was reversible when ZnSO4 was removed from the media. By analyzing v-myc mRNA levels, the inducible alterations in cellular morphology and growth were found to be associated with increased v-myc expression. The metallothionein promoter exhibited negligible constitutive expression of v-myc and none of the clones isolated exhibited spontaneous transformation. Our results show that the use of a metallothionein promoter v-myc construct facilitates the study of inducible fibroblast transformation.
Bonham
L
L
Children's Medical Research Foundation, Camperdown, Sydney, NSW, Australia.
Kwok
J
J
Chisholm
O
O
Symonds
G
G
D000818:Animals; D002472:Cell Transformation, Viral / Q000187:drug effects / Q000502:physiology*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D016259:Genes, myc / Q000502:physiology*; D008668:Metallothionein / Q000235:genetics / Q000502:physiology*; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D013431:Sulfates / Q000494:pharmacology; D014162:Transfection; D015032:Zinc / Q000494:pharmacology; D019287:Zinc Sulfate
D012333:RNA, Messenger; D013431:Sulfates; D019287:Zinc Sulfate; D008668:Metallothionein; D015032:Zinc