Hubé F, Reverdiau P, Iochmann S, Gruel Y
Inserm, U618, Tours, F-37032 France; IFR 135, Tours, F-37044 France.
Mol Biotechnol. 2005 Sep;31(1):81-4. doi: 10.1385/MB:31:1:081.
Most housekeeping genes, tumor-suppressor genes, and approx 40% of tissue-specific genes contain G+C sequences in their promoter region that were very difficult to amplify. In this report, we propose an improved polymerase chain reaction (PCR) method to be used for successful amplification of the tissue factor pathway inhibitor (TFPI)-2 gene promoter region that exhibit >70% G+C content in a sequence of approx 300 bp and a complete CpG island region spanning exon 1, the three transcription initiation sites, and the translation start site. Therefore, this method can be recommended to amplify other GC-rich genomic templates.
大多数管家基因、肿瘤抑制基因以及约40%的组织特异性基因在其启动子区域含有G+C序列，这些序列很难扩增。在本报告中，我们提出了一种改进的聚合酶链反应（PCR）方法，用于成功扩增组织因子途径抑制剂（TFPI）-2基因的启动子区域，该区域在一段约300 bp的序列中G+C含量>70%，并且包含一个完整的CpG岛区域，跨越外显子1、三个转录起始位点和翻译起始位点。因此，该方法可推荐用于扩增其他富含GC的基因组模板。
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