TY - JOUR
T1 - Induced Helicobacter pylori vacuolating cytotoxin VacA expression after initial colonisation of human gastric epithelial cells
AU - van Amsterdam, Karin
AU - van Vliet, Arnoud H. M.
AU - Kusters, Johannes G.
AU - Feller, Monique
AU - Dankert, Jacob
AU - van der Ende, Arie
PY - 2003
Y1 - 2003
N2 - To study the effect of initial colonisation on Helicobacter pylori gene expression, altered H. pylori gene transcription during co-culture with human gastric epithelial cells was determined. Therefore, an insertion library of H. pylori with random chromosomal fusions to a promoterless cat gene was grown in the presence of HM02 gastric epithelial cells and varying levels of chloramphenicol. One H. pylori transformant was chloramphenicol-resistant in the presence, but chloramphenicol-susceptible in the absence of gastric epithelial HM02 cells. This transformant had the promoterless cat gene inserted into the HP0887 gene, which encodes the vacuolating cytotoxin VacA, an important virulence factor of H. pylori. Reverse transcriptase polymerase chain reaction on cDNA of this transformant confirmed vacA upregulation near HM02 cells. These results show the applicability of this technique to study H. pylori gene regulation in its natural environment. (C) 2003 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved
AB - To study the effect of initial colonisation on Helicobacter pylori gene expression, altered H. pylori gene transcription during co-culture with human gastric epithelial cells was determined. Therefore, an insertion library of H. pylori with random chromosomal fusions to a promoterless cat gene was grown in the presence of HM02 gastric epithelial cells and varying levels of chloramphenicol. One H. pylori transformant was chloramphenicol-resistant in the presence, but chloramphenicol-susceptible in the absence of gastric epithelial HM02 cells. This transformant had the promoterless cat gene inserted into the HP0887 gene, which encodes the vacuolating cytotoxin VacA, an important virulence factor of H. pylori. Reverse transcriptase polymerase chain reaction on cDNA of this transformant confirmed vacA upregulation near HM02 cells. These results show the applicability of this technique to study H. pylori gene regulation in its natural environment. (C) 2003 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved
U2 - https://doi.org/10.1016/S0928-8244(03)00226-8
DO - https://doi.org/10.1016/S0928-8244(03)00226-8
M3 - Article
C2 - 14642310
SN - 0928-8244
VL - 39
SP - 251
EP - 256
JO - FEMS immunology and medical microbiology
JF - FEMS immunology and medical microbiology
IS - 3
ER -