Efficient transformation of human fibroblasts by adenovirus-simian virus 40 recombinants.
The origin-defective simian virus 40 (SV40) mutant 6-1 has been useful in transforming human cells (Small et al., Nature [London] 296:671-672, 1982; Nagata et al., Nature [London] 306:597-599, 1983). However, the low efficiency of transformation achieved by DNA transfection is a major drawback of the system. To increase the efficiency of SV40-induced transformation of human fibroblasts, we used recombinant adenovirus-SV40 virions which contain a complete SV40 early region including either a wild-type or defective (6-1) origin of replication. The SV40 DNA was cloned into the adenovirus vector in place of early region 1. Cell lines transformed by viruses containing a functional origin of replication produced free SV40 DNA. These cell lines were subcloned, and some of the subclones lost the ability to produce free viral DNA. Subclones that failed to produce free viral DNA were found to possess a mutated T antigen. Cell lines transformed by viruses containing origin-defective SV40 mutants did not produce any free DNA. Because of the high efficiency of transformation, we suggest that the origin-defective chimeric virus is a convenient system for establishing SV40-transformed cell lines from any human cell type that is susceptible to infection by adenovirus type 5.
D000260:Adenoviruses, Human / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D002472:Cell Transformation, Viral; D002678:Chimera; D003001:Cloning, Molecular; D003433:Crosses, Genetic; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000302:isolation & purification; D003673:Defective Viruses / Q000235:genetics; D005814:Genes, Viral; D006801:Humans; D009154:Mutation; D013539:Simian virus 40 / Q000235:genetics*
D004279:DNA, Viral; D004262:DNA Restriction Enzymes