High efficiency of replication and expression of foreign genes in SV40-transformed human fibroblasts.
Human fibroblasts (HF) were transformed in vitro with origin-defective SV40 DNA (ori-) using the calcium phosphate co-precipitation technique. The SV40 ori- transformed human cells (HSF) were able to replicate efficiently a recombinant DNA molecule containing the ori sequence of SV40 DNA. Transfection of HFS with pTBC1, a recombinant pi vx plasmid containing the herpes simplex virus thymidine kinase (HSV-TK) gene and the ori SV40 sequences, results in high levels of TK mRNA of correct size. The pTBC1 plasmid does not appear to contain 'poison' sequences and can be efficiently re-established in Escherichia coli after replication in human cells. This host vector system may be of great usefulness in studying the expression of human genes in human cells.
D000818:Animals; D000957:Antigens, Viral, Tumor / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D004261:DNA Replication; D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D005796:Genes; D005814:Genes, Viral; D006801:Humans; D010957:Plasmids; D013539:Simian virus 40 / Q000235:genetics* / Q000276:immunology; D018139:Simplexvirus / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection
D000957:Antigens, Viral, Tumor; D004274:DNA, Recombinant; D013937:Thymidine Kinase; D004262:DNA Restriction Enzymes