Preparation of E. coli RNA polymerase transcription elongation complexes by selective photoelution from magnetic beads.

ABSTRACT: In vitro studies of transcription frequently require the preparation of defined elongation complexes. Defined transcription elongation complexes (TECs) are typically prepared by constructing an artificial transcription bubble from synthetic oligonucleotides and RNA polymerase. This approach is optimal for diverse applications but is sensitive to nucleic acid length and sequence and is not compatible with systems where promoter-directed initiation or extensive transcription elongation is crucial. To complement scaffold-directed approaches for TEC assembly, I have developed a method for preparing promoter-initiated Escherichia coli TECs using a purification strategy called selective photoelution. This approach combines TEC-dependent sequestration of a biotin-triethylene glycol transcription

SUBMITTER: Strobel EJ 

PROVIDER: S-EPMC8212663 | biostudies-literature | 2021 Jul

REPOSITORIES: biostudies-literature