Efficient bacterial transcription of DNA nanocircle vectors with optimized single-stranded promoters.

ABSTRACT: We describe experiments aimed at establishing whether circular single-stranded DNAs can form promoters for bacterial transcription from small folded motifs. In vitro selection experiments were carried out on circular 103-nt DNA libraries encoding 40-nt randomized sequences as well as self-processing hammerhead ribozymes. Rounds of rolling circle transcription, reverse transcription-PCR, and recyclization were carried out to optimize transcription efficiency. Sequences were identified that are 80-fold more actively transcribed than the initial library by E. coli RNA polymerase (RNAP). The selected motifs were found to be more active than canonical E. coli promoters in the same context. Experiments also demonstrated that a single-stranded pseudopromoter identified by this selection can be tr

SUBMITTER: Ohmichi T 

PROVIDER: S-EPMC117513 | biostudies-literature | 2002 Jan

REPOSITORIES: biostudies-literature