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. 1995 Jun 16;81(6):925-34.
doi: 10.1016/0092-8674(95)90012-8.

Initiation of V(D)J recombination in a cell-free system

D C van Gent  1 , J F McBlane, D A Ramsden, M J Sadofsky, J E Hesse, M Gellert
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Free article

Initiation of V(D)J recombination in a cell-free system

D C van Gent et al. Cell. .
Free article

Abstract

Cells performing V(D)J recombination make specific cuts in DNA at recombination signal sequences. Here, we show that nuclear extracts of pre-B cell lines carry out this specific cleavage. The products of cleavage are the same as found previously in thymocytes: full-length, blunt, 5'-phosphorylated signal ends, and covalently sealed (hairpin) coding ends. A complete signal sequence is required. Recombinant RAG1 protein greatly increases activity and complements an inactive extract from a RAG1 (-/-) pre-B cell line. When the extracts are fractionated, cleavage activity correlates with the presence of RAG2 protein. These results suggest that RAG1 and RAG2 are components of the V(D)J recombinase.

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