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Nucleic Acids Research, 1989, Vol. 17, No. 10 3855-3863
© 1989

MOLECULAR BIOLOGY

Effect of the sequence-dependent structure of the 17 bp AT spacer on the strength of consensus-like E.coli promoters in vivo

Tomasz Lozinski, Wojciech T. Markiewicz1, Tadeusz K. Wyrzykiewicz1 and Kazimierz L. Wierzchowski*,

1Institute of Bioorganic Chemistry, Polish Academy of Sciences ul. Noskowskiego 12/14, Poznari, Poland Institute of Biochemistry and Biophysics, Polish Academy of Sciences ul. Rakowiecka 36, Warszawa 02-532

*To whom correspondence should be addressed.

Received February 16, 1989. Revised April 26, 1989. Accepted April 26, 1989.

Three E.coli promoters with the consensus sequences in the –35 and –10 regions and the 17 bp spacer made of random, heteronomous, and of both these classes of AT DNA simultaneously were constructed and cloned into plasmid pDS3. Electrophoretic gel mobilities of restriction fragments containing these promoters indicated that bending of the latter was proportional to the number of heteronomous AT DNA tracts. The strength of these promoters in vivo measured in relation to an internal transcriptional standard was shown to correlate well with gel mobilities of the respective restriction fragments and to decrease with the number of potential DNA bending sites encoded in the promoter structure.


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