A Nucleotide Element That Enhances Transcription from Bacterial Promoters
Inventors: Richard Gourse, Shawn Estrem, Tamas Gaal, Wilma Ross
The Wisconsin Alumni Research Foundation (WARF) is seeking commercial partners interested in developing a consensus sequence from the UP of E. coli that enhances transcription.
The main form of RNA polymerase in E. coli is Eσ70. One of the recognition elements within the promoter region for Eσ70 is the UP element. This element is situated upstream of the RNA transcription start site and acts as a transcription enhancer by binding to the α subunit of RNA polymerase.
UW-Madison researchers have obtained a series of 31 sequences for the UP element, each of which enhances transcription anywhere from 136- to 326-fold over the wild type sequence, and have derived a consensus sequence from these. The sequences were chosen based on how rapidly each bound to the bacterial RNA polymerase in a cell-free assay, assuming that the kinetics of the binding reaction determined sequence affinity for RNA polymerase.
- These sequences could improve the study of polymerase activation in a variety of bacterial systems
- The C-terminal domain of the alpha subunit of RNA polymerase, which interacts with the UP element, is highly conserved across many bacterial species, allowing the consensus sequence to be widely applicable.
- The large number of enhancing sequences available provides for nucleotide preferences in a number of bacterial systems.
- May be combined with core promoter elements recognized by most eukaryotic bacterial RNA polymerases
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