Harper Lucy V, Hilton Anthony C, Jones Alan F
Molecular Biosciences, School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham, B4 7ET, UK.
Mol Cell Probes. 2003 Oct;17(5):261-5. doi: 10.1016/s0890-8508(03)00063-x.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is an enzyme which catalyses the conversion of glyceraldehyde-3-phosphate to 1,3 diphosphoglycerate. It is considered to be constitutively expressed in all cells, and as such the gene for GAPDH (gapd) is commonly used as a benchmark reference in expression studies. However, previous investigations have demonstrated that gapd may show altered gene expression in a number of disease states and under certain experimental conditions, suggesting that results of experiments using gapd as a control should be interpreted with caution. Furthermore, consideration must be given to the potential co-amplification of pseudogenes of gapd during RT-PCR. Here, we describe a method to avoid the amplification of contaminating pseudogenes through the design of primers that bind only to genuine gapd mRNA transcript.
甘油醛-3-磷酸脱氢酶（GAPDH）是一种催化甘油醛-3-磷酸转化为1,3-二磷酸甘油酸的酶。它被认为在所有细胞中组成性表达，因此GAPDH基因（gapd）通常在表达研究中用作基准参考。然而，先前的研究表明，gapd在许多疾病状态和某些实验条件下可能表现出基因表达的改变，这表明以gapd作为对照的实验结果应谨慎解释。此外，在逆转录聚合酶链反应（RT-PCR）过程中，必须考虑gapd假基因的潜在共扩增。在此，我们描述了一种通过设计仅与真正的gapd mRNA转录本结合的引物来避免污染性假基因扩增的方法。
