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Cell. 1992 Apr 3;69(1):27-39.

P element transposition in vitro proceeds by a cut-and-paste mechanism and uses GTP as a cofactor.

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  • 1Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142.

Abstract

We have developed an in vitro reaction system for Drosophila P element transposition. Transposition products were recovered by selection in E. coli, and contained simple P element insertions flanked by 8 bp target site duplications as observed in vivo. Transposition required Mg+2 and partially purified P element transposase. Unlike other DNA rearrangement reactions, P element transposition in vitro used GTP as a cofactor; deoxyGTP, dideoxyGTP, or the nonhydrolyzable GTP analogs GMP-PNP or GMP-PCP were also used. Transposon DNA molecules cleaved at the P element termini were able to transpose, but those lacking 3'-hydroxyl groups were inactive. These biochemical data are consistent with genetic data suggesting that P element transposition occurs via a "cut-and-paste" mechanism.

PMID:
1313335
[PubMed - indexed for MEDLINE]
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