Tsutsumi H, Tani K, Fujii H, Miwa S
Department of Internal Medicine, University of Tokyo, Japan.
Genomics. 1988 Jan;2(1):86-9. doi: 10.1016/0888-7543(88)90112-7.
Pyruvate kinase (PK) has four isozymes (L, R, M1, M2) that are encoded by two different genes of PK L and M. Differential splicing produces L- and R-type PK mRNA and M1- and M2-type PK mRNA from the PK L gene and the PK M gene, respectively. The nucleotide sequences of the 3'-noncoding region are the same between the L- and the R-type PK and between the M1- and the M2-type PK. We isolated 3'-noncoding sequences of human L- and M2-type PK cDNA to construct L-type and M-type PK specific probes. With these probes, we performed Northern blot analysis of the RNA samples extracted from human tissues. Northern blot analysis showed that both kidney and liver had mRNAs that hybridized with both the L and M probes. Small intestine, skeletal muscle, brain, testis, and lung mRNAs hybridized only with the M probe. Our probes are considered useful for the detection of the types of PK isozymes expressed in small amounts, which are very difficult to detect using the conventional PK polyacrylamide gel electrophoretic method.
丙酮酸激酶（PK）有四种同工酶（L、R、M1、M2），它们由PK L和M的两个不同基因编码。差异剪接分别从PK L基因和PK M基因产生L型和R型PK mRNA以及M1型和M2型PK mRNA。L型和R型PK之间以及M1型和M2型PK之间3'-非编码区的核苷酸序列相同。我们分离了人L型和M2型PK cDNA的3'-非编码序列，以构建L型和M型PK特异性探针。利用这些探针，我们对从人体组织中提取的RNA样本进行了Northern印迹分析。Northern印迹分析表明，肾脏和肝脏都有与L和M探针都杂交的mRNA。小肠、骨骼肌、脑、睾丸和肺的mRNA仅与M探针杂交。我们的探针被认为可用于检测少量表达的PK同工酶类型，而使用传统的PK聚丙烯酰胺凝胶电泳方法很难检测到这些同工酶。
