TY - JOUR
T1 - Transposon-mediated site-specific recombination in vitro
T2 - DNA cleavage and protein-DNA linkage at the recombination site
AU - Reed, R. R.
AU - Grindley, N. D.F.
N1 - Funding Information:
We thank Martin Gellert, Kiyoshi Mizuuchi, Howard Nash, Charles Radcfing and Joan Steitz for stimulating discussions and helpful suggestions. We are also grateful to Arthur Landy, Cathy Joyce and Terry Platt for critical reading of the manuscript. This work was supported by grants from the National Institutes of Health.
PY - 1981/9
Y1 - 1981/9
N2 - Resolvase, the product of the tnpR gene of the transposable element γδ, mediates a site-specific recombination between two copies of the element directly repeated on the same replicon. The resolution site, res, at which resolvase acts lies in the intercistronic region between the tnpA and tnpR genes. We have studied this site-specific recombination in vitro. In the absence of Mg2+, a resolvase-res complex is formed, which contains DNA molecules that have been cleaved at res. Our data suggest that in this complex resolvase is covalently attached to the 5′ ends of the cleaved DNA, leaving free 3′ hydroxyl groups. DNA cleavage is stimulated by the interaction of two res sites on the same substrate molecule and appears to be an intermediate step in normal res site recombination. We show that the DNA is cut within a region previously identified as containing the crossover point at the palindromic sequence 5′- TT AT↓AA AA↑TA TT to generate 3′ extensions of two bases.
AB - Resolvase, the product of the tnpR gene of the transposable element γδ, mediates a site-specific recombination between two copies of the element directly repeated on the same replicon. The resolution site, res, at which resolvase acts lies in the intercistronic region between the tnpA and tnpR genes. We have studied this site-specific recombination in vitro. In the absence of Mg2+, a resolvase-res complex is formed, which contains DNA molecules that have been cleaved at res. Our data suggest that in this complex resolvase is covalently attached to the 5′ ends of the cleaved DNA, leaving free 3′ hydroxyl groups. DNA cleavage is stimulated by the interaction of two res sites on the same substrate molecule and appears to be an intermediate step in normal res site recombination. We show that the DNA is cut within a region previously identified as containing the crossover point at the palindromic sequence 5′- TT AT↓AA AA↑TA TT to generate 3′ extensions of two bases.
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U2 - 10.1016/0092-8674(81)90179-3
DO - 10.1016/0092-8674(81)90179-3
M3 - Article
C2 - 6269756
AN - SCOPUS:0019507482
SN - 0092-8674
VL - 25
SP - 721
EP - 728
JO - Cell
JF - Cell
IS - 3
ER -