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Isolation of transforming DNA: cloning the hamster aprt gene.
We have isolated the hamster gene coding for the enzyme adenine phosphoribosyl transferase (aprt) using gene transfer and molecular cloning of transforming DNA. Mouse aprt- cells were transformed to the aprt+ phenotype with the product of ligation of Hind III-cleaved hamster genomic DNA and pBR322 DNA. In this manner, the aprt gene was linked to a marked plasmid sequence and segregated from other hamster sequences. A lambda-recombinant phage containing pBR322 DNA sequences was isolated from a library of aprt+ transformed cell DNA. The phage DNA transfers hamster aprt+ activity at a frequency expected of a pure gene. Furthermore, sequences homologous to this clone are present in all hamster aprt+ transformants examined. This experimental design should in theory permit the isolation of any gene coding for selectable or identifiable functions for which DNA-mediated gene transfer can be effected.[1]References
- Isolation of transforming DNA: cloning the hamster aprt gene. Lowy, I., Pellicer, A., Jackson, J.F., Sim, G.K., Silverstein, S., Axel, R. Cell (1980) [Pubmed]
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