Toniolo D, Persico M G, Battistuzzi G, Luzzatto L
Mol Biol Med. 1984 Apr;2(2):89-103.
Glucose-6-phosphate dehydrogenase (G6PD) is a household enzyme that accounts, in many cells, for about 0.03% of cellular protein. We have developed an assay for G6PD-specific mRNA based on in vitro translation of RNA from human fibroblasts and immunoprecipitation of the translation products with an anti-G6PD antiserum. By making use of this assay, G6PD mRNA has been purified 50 to 100-fold in three steps. We estimate that the mRNA encoding G6PD constitutes less than 0.02% of total poly(A)+ RNA in human fibroblasts. The size of the G6PD mRNA has been established in denaturing conditions as being in the range between 2800 and 3200 nucleotides. This has been confirmed by Northern blot analysis. Since the G6PD coding sequence is estimated to be about 1491 nucleotides, the G6PD mRNA has long untranslated sequences, most of which is at the 3' end and which may be heterogeneous in length. The sequence of the last 608 nucleotides of this mRNA has been determined.
葡萄糖-6-磷酸脱氢酶（G6PD）是一种常见的酶，在许多细胞中约占细胞蛋白质的0.03%。我们基于人成纤维细胞RNA的体外翻译以及用抗G6PD抗血清对翻译产物进行免疫沉淀，开发了一种检测G6PD特异性mRNA的方法。利用该方法，G6PD mRNA已通过三个步骤纯化了50至100倍。我们估计，编码G6PD的mRNA在人成纤维细胞中占总poly(A)+ RNA的比例不到0.02%。在变性条件下确定G6PD mRNA的大小在2800至3200个核苷酸之间。这已通过Northern印迹分析得到证实。由于G6PD编码序列估计约为1491个核苷酸，G6PD mRNA具有长的非翻译序列，其中大部分在3'端，且长度可能不均一。已确定该mRNA最后608个核苷酸的序列。
