Real-time quantitative PCR analysis of viral transcription.
Real-time quantitative PCR analysis of viral transcription.
复制标题
病毒转录的实时定量 PCR 分析。
作者：
James F. Papin;Wolfgang Vahrson;Rebecca Hines;D. Dittmer
使用 DNA 阵列进行全基因组分析使我们对细胞生物学的理解取得了巨大进步。当应用于大型病毒基因组（例如疱疹病毒）时，它也取得了类似的成功。不幸的是，大多数 DNA 阵列仍然需要专门且昂贵的资源，并且通常需要大量的输入 RNA。另一种方法是使用实时定量 PCR 查询整个病毒基因组。我们为人类疱疹病毒 8 的每个开放阅读框设计了这种基于 PCR 的阵列，并在此描述了量化病毒 mRNA 的一般设计标准、验证程序和详细应用。这应该为全基因组阵列或仅测量任何一种特定感兴趣的 mRNA 的转录提供有用的资源。由于这些芯片是基于 RT-PCR 的，因此它们本质上比当前基于杂交的方法更灵敏、更稳健，非常适合在发病机制模型中查询病毒基因表达。
Whole-genome profiling using DNA arrays has led to tremendous advances in our understanding of cell biology. It has had similar success when applied to large viral genomes, such as the herpesviruses. Unfortunately, most DNA arrays still require specialized and expensive resources and, generally, large amounts of input RNA. An alternative approach is to query entire viral genomes using real-time quantitative PCR. We have designed such PCR-based arrays for every open reading frame of human herpesvirus 8 and describe here the general design criteria, validation procedures, and detailed application to quantify viral mRNAs. This should provide a useful resource either for whole-genome arrays or just to measure transcription of any one particular mRNA of interest. Because these arrays are RT-PCR-based, they are inherently more sensitive and robust than current hybridization-based approaches and are ideally suited to query viral gene expression in models of pathogenesis.
影响因子：
7.5
作者：
Bentley, D
通讯作者：
Bentley, D