PbrSGRL DAP seq

To investigate the target binding elements of PbrSGRL, we employed Red Zaosu fruit peel as material for library construction. Genomic DNA was extracted from Red Zaosu samples, fragmented, and ligated with Illumina based sequencing adapters to construct a genomic DNA library. In vitro expression of PbrSGRL protein: The coding sequence (CDS) for PbrSGRL was cloned into a vector bearing an affinity tag for in vitro protein expression, yielding a fusion protein comprising the transcription factor and affinity tag. Affinity purification and sequencing: The purified PbrSGRL affinity tag fusion protein was incubated with the genomic DNA library. The target protein-DNA complex was extracted and purified using Halo Tag specific magnetic beads. Captured DNA fragments were amplified via PCR and sequenced using the Illumina sequencing platform. The reference genome information is NCBI RefSeq assembly GCF 000315295.1 (Submitted GenBank assembly GCA 000315295.1), which provides a basis for PbrSGRL target gene screening based on the sequencing results.