Acid-induced gene expression in Helicobacter pylori: study in genomic scale by microarray.
To understand the RNA expression in response to acid stress of Helicobacter pylori in genomic scale, a microarray membrane containing 1,534 open reading frames (ORFs) from strain 26695 was used. Total RNAs of H. pylori under growth conditions of pH 7.2 and 5.5 were extracted, reverse transcribed into cDNA, and labeled with biotin. Each microarray membrane was hybridized with cDNA probe from the same strain under two different pH conditions and developed by a catalyzed reporter deposition method. Gene expression of all ORFs was measured by densitometry. Among the 1,534 ORFs, 53 ORFs were highly expressed (> or = 30% of rRNA control in densitometry ratios). There were 445 ORFs which were stably expressed (<30% of rRNA in densitometry) under both pH conditions without significant variation. A total of 80 ORFs had significantly increased expression levels at low pH, while expressions of 4 ORFs were suppressed under acidic condition. The remaining 952 ORFs were not detectable under either pH condition. These data were highly reproducible and comparable to those obtained by the RNA slot blot method. Our results suggest that microarray can be used in monitoring prokaryotic gene expression in genomic scale.
Ang
S
S
Graduate Institute of Microbiology, National Taiwan University College of Medicine, Taipei, Taiwan.
Lee
C Z
CZ
Peck
K
K
Sindici
M
M
Matrubutham
U
U
Gleeson
M A
MA
Wang
J T
JT
D000143:Acids / Q000494:pharmacology*; D000220:Adaptation, Biological / Q000235:genetics*; D015536:Down-Regulation; D020869:Gene Expression Profiling / Q000379:methods*; D016680:Genome, Bacterial; D016480:Helicobacter pylori / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016366:Open Reading Frames; D012329:RNA, Bacterial / Q000302:isolation & purification; D012333:RNA, Messenger / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D015854:Up-Regulation
D000143:Acids; D012329:RNA, Bacterial; D012333:RNA, Messenger