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Science 20 October 1989:
Vol. 246. no. 4928, pp. 358 - 363
DOI: 10.1126/science.2678475
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Articles
Science, Vol 246, Issue 4928, 358-363
Copyright © 1989 by American Association for the Advancement of Science
Stabilization of Z DNA in vivo by localized supercoiling
AR Rahmouni
and
RD Wells
Department of Biochemistry, School of Medicine, University of Alabama, Birmingham 35294.
Biological processes such as transcription may generate domains of supercoiling on a circular DNA. The existence of these domains in Escherichia coli was investigated by the ability of different lengths of (CG) tracts, cloned upstream or downstream from the tetracycline resistance gene (tet) of pBR322, to adopt the Z structure in vivo. Segments as short as 12 base pairs adopt the Z form when cloned upstream from the tet gene (Eco RI site), whereas no Z DNA was detected when this sequence was cloned downstream (Sty I site), even with a 74-base pair (CG) tract that requires less supercoiling than shorter tracts for the B-Z transition. Hence the localized supercoil density in pBR322 can be as high as -0.038 and as low as -0.021 at different loci. These data demonstrate the existence of the Z structure for commonly found natural sequences and support the notion of domains of negative supercoiling in vivo.
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