Identify the binding sites of mTERF5 in chloroplast of Arabidopsis thaliana through cpChIP-seq (additional experiments)

We performed chloroplast ChIP-seq (cpChIP-seq) to identify the possible DNA-binding sites of mTERF5 in Arabidopsis thaliana. To this end, we generated transgenic Arabidopsis plants expressing mTERF5 carrying an HA tag under the control of the CaMV 35S promoter. Then, We used the polyclonal antibody (abcam, ab9110, lot GR304617-8 ) against HA tag which conjugated to ChIP-Grade protein A/G agarose (Thermo scientific, 26161, lot QJ223903) to perform cpChIP assay. The obtained chromatin immunoprecipitated DNA of chloroplasts were used to build DNA libaries for high-throughput sequencing. Finally, we showed that three potenssial DNA regions across the chloroplast genome compared to the control group were enriched by mTERF5. Overall design: Deep sequencing was used for examination of mTERF5 associated DNA regions in chloroplast of Arabisopsis thaliana. Two biological replicates were performed, and the "Input sample" was used as a reference sample. Rabbit IgG (abmart, B30011, lot 294670) was used as negetive antibody control for IP.