Abstract
The direct approach of capillary microinjection was used to study exogenous gene expression in mouse and human cells. The efficiencies of biochemical and morphological transformation in mouse cells and the differential ras gene expression in mouse and human cells were determined. Biochemical transformation efficiency of mouse cells after microinjection was ≈2.5% with the thymidine-kinase plasmid and ≈30% with the neomycin-resistant gene, pSV2 neo, which contains the SV-40 enhancer sequence. Mouse NIH3T3 cells microinjected with genes of the ras family—including viral isolates, the human bladder oncogene EJ, and N- ras—produced morphologically transformed foci with an efficiency of 10–20%, whereas the normal human cellular homologue of the ras gene in Harvey sarcoma virus showed an efficiency of 0.5–1%. Expression of v- ras genes was compared in mouse and human cells by both immunofluorescence detection of the ras protein, p21, 24 hr after microinjection and by the focus assay. The data suggest that microinjection is a useful approach for the investigation of early exogenous gene expression and control in eukaryotic cells and has the potential to allow insight into oncogenic events.
Published Version
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