Nuclei isolated from a permanent cell line derived from Drosophila melanogaster embryos have been injected, along with a radioactive DNA precursor [3H]TTP, into Xenopus laevis eggs. In culture, less than 7% of the cells were in S phase. After a 90 min incubation, following injection into eggs, 99% of the nuclei were shown by autoradiography to have synthesized DNA. In a similar experiment, a density label BrdUTP was injected into eggs along with the nuclei. Subsequent analysis on caesium chloride gradients showed that this DNA synthesis was semi-conservative replication. Therefore we conclude that signals present in Xenopus egg cytoplasm can initiate and sustain true semi-conservative DNA replication in nuclei from an invertebrate organism.