Transcriptome analysis of Vibrio vulnificus cells that are either exposed to HT29-methotrexate intestinal epithelial cells or grown in basal medium eagle

ABSTRACT: The pathogenic marine bacterium Vibrio vulnificus is a causative agent of food-borne diseases such as life-threatening septicemia and gastroenteritis in individuals with underlying predisposed conditions.Because most infection cases of V. vulnificus are caused by ingestion of contaminated seafood, the pathogen has to encounter host cells in the gut at the early stage of infection and alters its gene expression patters to adapt the gut environment of host. To identify differentially expressed genes when V. vulnificus cells are exposed to host cells, a transcriptome analysis of V. vulnificus cells that were either exposed to HT29-methotrexate intestinal epithelial cells or grown in basal medium eagle (Sigma) (as a negative control) was performed using Illumina GAIIx system. 16S and 23S rRNA were depleted using MICROBExpress kit (Ambion) from the total RNAs of each sample. Strand-specific cDNA libraries of the samples were constructed using Truseq stranded mRNA prep. kit (Illumina). Single-end and 36 cycle sequencing were performed using Iluumina GAIIx sequencer by ChunLab (http://www.chunlab.com) and Raw sequence data were analyzed by CLC Genomics Workbench 5. 5. 1 (CLC Bio)

INSTRUMENT(S): PCR machine; Superscript II polymerase Kit (Invitrogen); PCR machine; AMPure XP beads; TruSeq stranded mRNA sample prep kit.; TruSeq Stranded mRNA stranded prep. kit, Centrifuge 5424R (Eppendorf); miRNeasy Mini Kit (Qiagen); TURBO^TM DNase (Ambion); Bioanalyser (Agilent), Centrifuge 5424R (Eppendorf); RNAprotect Bacteria Reagent (Qiagen), PCR machine; AMPure XP beads; TruSeq stranded mRNA sample prep. kit., PCR machine; AMPure XP beads; TruSeq Stranded mRNA stranded prep. kit, Centrifuge 5424R (Eppendorf); RNAprotect Bacteria Reagent (Qiagen); Cell culture dish (Nunc), Illumina Genome Analyzer IIx, PCR machine; MICROBExpress kit (Ambion); TruSeq Stranded mRNA stranded prep. kit

ORGANISM(S): Vibrio vulnificus

SUBMITTER: Kyung Ku Jang 

PROVIDER: E-MTAB-5629 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress