Interaction of Escherichia coli RNA Polymerase with Artificial Promoters, Containing Nonnucleotide Spacers

Abstract

To study the functional role of the spacer region between two consensus −10 and −35 elements of promoters, recognized by E. coli RNA polymerase, the model promoter-like DNA duplexes containing nonnucleotide inserts (mimicking 17-mer spacer) either in one or both strands, were constructed. The modified duplexes can form the heparin-resistant binary complexes with RNA polymerase. The DNA duplex with nonnucleotide insert in the template strand can specifically direct the synthesis of mRNA in the in vitro run-off transcription assays.

Keywords:

• E. coli RNA polymerase
• promoters
• modified oligonucleotides