ChIP-sequencing using anti-BldD antibody, vegetative growth in Actinoplanes missouriensis

ChIP-sequencing was conducted using anti-BldD antibody in the rare actinomycete Actinoplanes missouriensis to identify binding regions of DNA-binding protein BldD in vegetative growth. The bacterium forms motile zoospore-containing sporangia upon the substrate mycelia under adverse conditions. In this project, A. missouriensis was cultured in PYM nutrient-rich liquid culture for 2 days. The culture was subjected to formaldehyde crosslinking and sonication. After immunoprecipitation using new generated anti-BldD antibody and reverse crosslinking, sequencing libraries were prepared from the obtained DNA (ChIP DNA) together with a total DNA control (Input DNA) and sequenced with a MiSeq sequencer.