Weiskirchen R, Siemeister G, Hartl M, Bister K
Institute of Biochemistry, School of Medicine, University of Cologne, Germany.
Gene. 1993 Jun 30;128(2):269-72. doi: 10.1016/0378-1119(93)90573-l.
Using differential hybridization techniques, a cDNA library derived from a line of v-myc-transformed quail embryo fibroblasts was screened for clones whose expression was elevated in transformed, as compared with normal, cells. One of the isolated clones contained the entire coding region of the quail glyceraldehyde-3-phosphate dehydrogenase-encoding gene (GAPDH). A comparison of the deduced 333-amino-acid (aa) sequence of quail GAPDH with that of the only other avian (chicken) GAPDH sequence known, and with those of mammalian counterparts indicates the strong aa sequence conservation of this glycolytic enzyme. GAPDH is expressed in all transformed and non-transformed quail and chicken embryo fibroblasts and macrophages tested, with a moderate elevation of expression in most of the transformed cell lines. In the avian genomes, GAPDH is present in a single copy, in contrast to the high number of GAPDH-related sequences in mammalian species.
利用差异杂交技术,对源自v-myc转化的鹌鹑胚胎成纤维细胞系的cDNA文库进行筛选,以寻找那些在转化细胞中表达相较于正常细胞有所升高的克隆。分离得到的一个克隆包含鹌鹑甘油醛-3-磷酸脱氢酶编码基因(GAPDH)的完整编码区。将推导的鹌鹑GAPDH的333个氨基酸序列与唯一已知的另一种禽类(鸡)GAPDH序列以及哺乳动物对应序列进行比较,表明这种糖酵解酶的氨基酸序列具有很强的保守性。在所测试的所有转化和未转化的鹌鹑及鸡胚胎成纤维细胞和巨噬细胞中均有GAPDH表达,在大多数转化细胞系中表达有适度升高。与哺乳动物物种中大量的GAPDH相关序列不同,在禽类基因组中,GAPDH以单拷贝形式存在。