Abstract
A series of replication-defective retroviral vectors were assessed for their ability to efficiently transfer functional genes into undifferentiated cells. In these vectors (designated handicapped because of a deletion of enhancer and promoter sequences in the viral long terminal repeat) transcription of inserted genes is under the control of internal promoters. Although a composite promoter composed of a mutant polyoma virus enhancer (PyF441) coupled to the herpes simplex virus thymidine kinase promoter was anticipated to function efficiently, it was found to be significantly inferior to the mouse X-chromosome phosphoglycerate kinase ( pgk-1) promoter, in its ability to express the selectable neomycin phosphotransferase gene in mouse embryonal carcinoma cells. The pHMB vector, which contains the pgk-1 promoter, was shown to confer the drug-resistant phenotype at high frequencies to F9 and P19 cells. This vector might prove to be of general utility for efficient gene expression in other developmental contexts.
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