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Nucleic Acids Research, 1990, Vol. 18, No. 20 5961-5967
© 1990

MOLECULAR BIOLOGY

Selection for mutations in the PR promoter of bacteriophage lambda

Susan Brown1, Julia Ferm1, Scott Woody1,2 and Gary Gussin1,2

1Department of Biology, University of Iowa Iowa City IA 52242, USA 2Genetics PhD Program, University of Iowa Iowa City IA 52242, USA

Received July 31, 1990. Revised September 14, 1990. Accepted September 14, 1990.

Insertion of DNA containing PR the early rlghtward promoter of bacteriophage lambda, is lethal to M13-derived vectors when the promoter directs transcription (using the ‘+’ strand as template) toward the M13 origin of replication (ori). Lethality can be relieved by mutation of PR, repression of the promoter by the {lambda} cl repressor, or by insertion of a strong transcription terminator between PR and ori. We have used selection for plaque formation in the absence of repressor to isolate 14 different mutations at 8 sites in PR. This method of isolating promoter mutants in vivo is applicable generally to strong promoters whose activity is regulated either positively or negatively.


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