Singh V K, Chader G J, Rodriguez I R
Laboratory of Retinal Cell and Molecular Biology, National Eye Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Mol Vis. 1998 Apr 20;4:7.
Pigment epithelium-derived factor (PEDF), a neurotrophic protein, is a member of the serine protease inhibitor supergene family. PEDF promotes both neuronal phenotype in cultured Y79 cells and neuronal survival in cultured cerebellar granulocytes. The purpose of this study was to clone the mouse PEDF gene and to determine its structure and levels of expression in different tissues.
The mouse PEDF cDNA was cloned from a mouse liver cDNA library using human PEDF cDNA as a probe. The mouse PEDF gene was cloned from a mouse ES genomic P1 library. DNA sequencing was performed using a PE-Applied Biosystems model 373 automated fluorescent sequencer.
The mouse PEDF cDNA is 1461 bp in length and contains an open reading frame of 417 amino acids. The mouse PEDF gene spans approximately 13 kb and, like the human, it is fragmented into 8 exons. The splice sites follow the AG/GT consensus rule. Southern blot analysis indicates that the mouse genome contains only one gene for PEDF. Northern blot analysis shows the presence of the PEDF transcript in a broad range of adult mouse tissues with liver showing the highest level of expression.
The mouse and human PEDF promoters share overall 27% similarity but are nearly identical between mouse +86 to -166 and human +102 to -96. The present study will allow us to move from in vitro experiments to in vivo studies through the development of a "knock-out" mouse model.
色素上皮衍生因子（PEDF）是一种神经营养蛋白，属于丝氨酸蛋白酶抑制剂超基因家族成员。PEDF可促进培养的Y79细胞的神经元表型以及培养的小脑粒细胞的神经元存活。本研究的目的是克隆小鼠PEDF基因，并确定其在不同组织中的结构和表达水平。
以人PEDF cDNA为探针，从小鼠肝脏cDNA文库中克隆小鼠PEDF cDNA。从小鼠胚胎干细胞基因组P1文库中克隆小鼠PEDF基因。使用PE-Applied Biosystems 373型自动荧光测序仪进行DNA测序。
小鼠PEDF cDNA长度为1461 bp，包含一个417个氨基酸的开放阅读框。小鼠PEDF基因跨度约13 kb，与人的一样，它被分割为8个外显子。剪接位点遵循AG/GT一致规则。Southern印迹分析表明，小鼠基因组中仅含有一个PEDF基因。Northern印迹分析显示，在成年小鼠的多种组织中均存在PEDF转录本，其中肝脏中的表达水平最高。
小鼠和人PEDF启动子总体上有27%的相似性，但在小鼠+86至-166和人+102至-96之间几乎相同。本研究将使我们能够通过开发“基因敲除”小鼠模型，从体外实验转向体内研究。
