Analysis of gene expression in Treponema denticola with differential display polymerase chain reaction.
Analysis of gene expression in Treponema denticola with differential display polymerase chain reaction.
复制标题
用差异显示聚合酶链反应分析齿垢密螺旋体的基因表达。
DOI：
10.1034/j.1399-302x.2000.150506.x
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发表时间：
2000
影响因子：
--
通讯作者：
Shi,W
中科院分区：
文献类型：
--
作者：
Tsai,JP;Shi,W
齿状密螺旋体是一种与牙周病相关的口腔螺旋体。 T 的大量分子成分。由于牙齿菌的基因组测序计划正在进行中，很快就会被人们所了解。基因组测序之后最重要的工作之一就是分析这些基因的功能及其调控。然而，与许多其他口腔病原体一样，目前可用于研究 T 基因表达的分子和遗传工具数量非常有限。齿垢。在本文中，我们描述了一种将差异显示聚合酶链式反应 (ddPCR) 用于 T 的方法。齿状系统。为了测试该方案的有效性，我们使用了三种不同的温度条件：4°C、25°C 和 42°C，来测试差异基因表达。在不同的 ddPCR 条件下，我们发现了许多差异表达的基因。其中一些差异表达基因被克隆并测序，发现与已知的温度调节基因（包括 HtrA）同源。研究表明ddPCR方法可以有效地应用于T. denticola用于分析各种条件下的基因表达。
Treponema denticolais an oral spirochete associated with the periodontal diseases. A great deal of the molecular components ofT. denticolawill be learned soon since its genome sequence project is on the way. One of the most important works after genome sequence is to analyze the function of these genes and their regulation. However, like many other oral pathogens, there are currently a very limited number of molecular and genetic tools available to study gene expression inT. denticola. In this article, we describe a method of adapting differential display polymerase chain reaction (ddPCR) for use in theT. denticolasystem. To test for effectiveness of this protocol, we used three different temperature conditions, 4°C, 25°C and 42°C, to test for differential gene expression. With various ddPCR conditions, we found a number of genes that were expressed differentially. Some of these differentially expressed genes were cloned and sequenced and found to be homologous with the known temperature‐regulated genes, including HtrA. The study indicates that the ddPCR method can be effectively used inT. denticolafor analyzing gene expression under various conditions.