Genome-wide characterization of cis-acting DNA targets of Opaque11 in maize

Opaque11 (O11) is a bHLH transcription factor that plays important roles in maize endosperm development, nutrient metabolism, and stress response. Overall design: Examination of 15 days after pollination (DAP) wild type maize endosperm with O11 specific antibody. o11 mutant seeds which have no O11 protein were used as negtive control. Please note that antibody for O11 was custom-made as following. Full-length O11 cDNA was cloned into the EcoR I and Xho I sites of the pET-32a vector (Novagen) and transformed into BL21. The 6xHis fusion recombinant O11 protein was purfied with the KTA purfication system using a 1 mL HisTrap FF crude column (GE Healthcare). Antibodies were prepared in rabbits by Shanghai ABclonal Biotechnology.