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Acid-Induced Gene Expression in Helicobacter pylori: Study in Genomic Scale by Microarray

机译：酸诱导幽门螺杆菌的基因表达：基因组规模的芯片研究。

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摘要

To understand the RNA expression in response to acid stress ofHelicobacter 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 (≧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.
机译：为了了解在基因组规模上幽门螺杆菌对酸胁迫的响应中的RNA表达，使用了包含来自26695株的1,534个开放阅读框（ORF）的微阵列膜。 H的总RNA。提取在pH 7.2和5.5的生长条件下的幽门螺杆菌，反转录成cDNA，并用生物素标记。将每个微阵列膜与来自相同菌株的cDNA探针在两个不同的pH条件下杂交，并通过催化的报道分子沉积方法进行显影。通过密度测定法测量所有ORF的基因表达。在1,534个ORF中，有53个ORF被高度表达（在光密度测定比率中，rRNA对照的≥30％）。在两个pH条件下，有445个ORF稳定表达（光密度法中rRNA的<30％），且无明显变化。在酸性条件下，总共80个ORF的表达水平显着增加，而4个ORF的表达被抑制。在任一pH条件下均无法检测到其余952个ORF。这些数据具有很高的可重复性，并且与通过RNA狭缝印迹法获得的数据相当。我们的结果表明，微阵列可用于监测基因组规模的原核基因表达。

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