Transcription profiling of Streptococcus pneumoniae after incubation with host respiratory tract epithelial cells

ABSTRACT: In order to study pneumococcal gene expression changes after incubation with host epithelial cells, logarithmic-phase grown S. pneumoniae strains (82 and R6) were incubated with host epithelial cells (A549 and ETEC) in T75 cell culture flasks at an MOI range of 10:1 ~ 50:1. After incubation for 1 h or 3 h, bacterial solution was removed and 7 ml of host cell lysis buffer which can remove host cells and stablize bacterial RNA was added immediately. Bacteria incubated with antibiotic-free media (MEM or DMEM) with 1% (v/v) FBS for the same time periods were used as medium controls. The RNA samples were isolated with RiboPure Bacteria kit (Ambion) or RNeasy Minikit (Qiagen), and purified with DNase I treatment. cDNA samples were synthesized and labeled with biotin-dUTP (samples) and fluorescei

ORGANISM(S): Streptococcus pneumoniae

SUBMITTER: Xin-Ming Song 

PROVIDER: E-TABM-187 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress