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Journal of Virology, July 2000, p. 6213-6216, Vol. 74, No. 13
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

DNA Sequence Motifs Which Direct Adeno-Associated Virus Site-Specific Integration in a Model System

Patricio Meneses,1 Kenneth I. Berns,2,* and Ernest Winocour3

Department of Pathology, Harvard Medical School, Boston, Massachusetts 022151; University of Florida College of Medicine, Gainesville, Florida 32610-00142; and Department of Molecular Genetics, Weizmann Institute of Science, Rehovot 76100, Israel3

Received 14 February 2000/Accepted 25 March 2000

The DNA sequence motifs which direct adeno-associated virus type 2 site-specific integration are being investigated using a shuttle vector, propagated as a stable episome in cultured cell lines, as the target for integration. Previously, we reported that the minimum episomal targeting elements comprise a 16-bp binding motif (Rep binding site [RBS]) for a viral regulatory protein (Rep) separated by a short DNA spacer from a sequence (terminal resolution site [TRS]) that can serve as a substrate for Rep-mediated nicking activity (R. M. Linden, P. Ward, C. Giraud, E. Winocour, and K. I. Berns, Proc. Natl. Acad. Sci. USA 93:11288-11294, 1996; R. M. Linden, E. Winocour, and K. I. Berns, Proc. Natl. Acad. Sci. USA 93:7966-7972, 1996). We now report that episomal integration depends upon both the sequence and the position of the spacer DNA separating the RBS and TRS motifs. The spacer thus constitutes a third element required for site-specific episomal integration.

* Corresponding author. Mailing address: University of Florida College of Medicine, 1600 SW Archer Rd., Room H-102, P.O. Box 100014, Gainesville, FL 32610-0014. Phone: (352) 392-2761. Fax: (352) 392-9395. E-mail: kberns{at}vpha.ufl.edu.

Journal of Virology, July 2000, p. 6213-6216, Vol. 74, No. 13
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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