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Study of recombination activity in mammalian cell extracts
- B. P. Konstantinov Institute of Nuclear Physics, Academy of Sciences of the USSR
Gatchina, Leningrad distr., USSR
The cell-free system utilizing nuclear extracts from human mieloblasts, HeLa cells and rat testes to initiate recombination events between homologous plasmids containing different mutant alleles of the Tc gene has been described. The incubation of the plasmids with nuclear extracts increased the frequency of Tcr recombinants from 2.6·105 to 2.7·10-3. The reaction required Mg2+ and was ATP- and dNTP-independent. In addition while using a sensitive homologous recombination strand-transfer assay ATP-independent strand-transfer activity from nuclear extracts of rat testis have been partially purified. It was homology-dependent and required the presence of Mg2+. Similarities between recombinase activity and other known eukaryotic recombinases are under discussion.
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