Abstract
The Escherichia coli (E. coli) Lac repressor is one of the best characterized sequence specific DNA binding proteins. The chapter discusses the mode of action of the repressor-operator complex on the transcription elongation complex (TEC) analyzed lac operon transcripts in vivo and transcripts generated from operator containing constructs in vivo and in vitro. Several studies have shown that the lac repressor is a versatile transcriptional roadblock in vivo and in vitro. Roadblocked TECs remain active and can be probed by a number of physical, chemical, and enzymatic methods. The exact state of the halted TECs depends on the sequences in the vicinity of the halt site. The chapter discusses effect of a lac repressor roadblock on expression of a downstream gene in vivo and transcription in vitro. The protocol for roadblocking the TEC with lac repressor is given.
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