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Gene, 18 (1982) 247-255 Elsevier Biomedical Press
 

Summary: Gene, 18 (1982) 247-255
Elsevier Biomedical Press
241
Specific protection of nucleotides in the lac operator from DMS
methylation and DNase I nicking by crude bacterial cell extracts
(Escherichia coli; DNA footprinting; regulatory proteins)
Gary B. Ruvkun, Venkatesan Sundaresau and Frederick M. Ausubel
Cellular and Developmental Group, Department of Biology, Harvard University, 16 Divinity Avenue, Cambridge, MA
02138 (U.S.A.)
(Received January 26th, 1982)
(Accepted March 22nd, 1982)
SUMMARY
Crude bacterial cell extracts prepared from an Escherichia coli fucIq strain were shown to protect specific
nucleotides in the fuc operator from methylation by dimethyl sulfate (DMS) or digestion by DNase I,
whereas no protection was observed using extracts prepared from a nearly isogenic lucZ_ strain. These
experiments show that it is not necessary to use purified regulatory proteins in experiments designed to
localize sequences on DNA which interact with proteins. Therefore, crude cell extracts should be useful in
DNA "footprinting" experiments to define regions of DNA which bind to unknown regulatory proteins.
INTRODUCTION
Transcriptional control in prokaryotes is gener-

  

Source: Ausubel, Frederick M. - Department of Genetics, Harvard University

 

Collections: Biology and Medicine