Building an improved transcription factor-centered yeast one hybrid system to identify DNA motifs bound by protein comprehensively.

ABSTRACT:

Background

Identification of the motifs bound by a transcription factor (TF) is important to reveal the function of TF. Previously, we built a transcription factor centered yeast one hybrid (TF-Centered Y1H) that could identify the motifs bound by a target TF. However, that method was difficult to comprehensively identify all the motifs bound by a TF.

Results

Here, we build an improved TF-Centered Y1H to comprehensively determine the motifs bound by a target TF. Recombination-mediated cloning in yeast was performed to construct a saturated prey library that contains 7 random base insertions. After TF-Centered Y1H screening, all the positive clones were pooled together to isolate pHIS2 vector. The insertion regions of pHIS2 were PCR amplified and the PCR product was subjected

SUBMITTER: Jingwen W 

PROVIDER: S-EPMC10158250 | biostudies-literature | 2023 May

REPOSITORIES: biostudies-literature