Gene expression of Acanthamoeba castellanii under long-term co-culture condition

The gene expression of A. castellanii cultured with low-nutrient medium and heat-killed bacteria for 21 days. The RNA-seq analysis was performed by Illumina HiSeq platform. The treated A. castellanii was compared with amoeba cultured in growth medium. Overall design: The A. castellanii as cultured with 20% PYG medium was diluted 1:4 by Page's Amoeba Saline (PAS) and heat-killed E. coli. To produce the heat-killed E. coli DH5a, the bacteria with OD = 1.0 was collected by centrifugation and suspended in the corresponding medium (100% or 20% PYG medium). Then, the suspended E. coli DH5a were heated at 100? for 10 minutes. The heat-killed bacteria were finally diluted into 2.0×106 CFU/mL. Hundred units/ml of ampicillin were added to each medium.