Fig. 1

The principle and procedure of the improved Transcription Factor-Centered yeast one hybrid (TF-Centered Y1H). A 7 bp random DNA sequence was inserted into pHIS2 at the digestion site of Sma I to form a library using the yeast recombination method, which was served as prey. The studied TF was cloned into pGADT7-Rec2 and was used as bait. After Y1H screening, all the positive clones were harvested and pooled together for pHIS2 isolation. The truncated pHIS2 containing the insertion sequence (125 bp in length) was PCR amplified, and the PCR products were analyzed using high-throughput sequencing. The insertion sequences were retrieved from the reads. The reads were further analyzed using MEME or other programs to find the conserved DNA motifs, which are potentially bound by the aim TF