MAMMALIAN-CELL LINES CAN BE EFFICIENTLY ESTABLISHED INVITRO UPON EXPRESSION OF THE SV40 LARGE T-ANTIGEN DRIVEN BY A PROMOTER SEQUENCE DERIVED FROM THE HUMAN VIMENTIN GENE
MAMMALIAN-CELL LINES CAN BE EFFICIENTLY ESTABLISHED INVITRO UPON EXPRESSION OF THE SV40 LARGE T-ANTIGEN DRIVEN BY A PROMOTER SEQUENCE DERIVED FROM THE HUMAN VIMENTIN GENE
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DOI：
10.1016/0248-4900(91)90003-6
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发表时间：
1991-01-01
影响因子：
2.7
通讯作者：
PAULIN, D
中科院分区：
文献类型：
--
作者：
SCHWARTZ, B;VICART, P;PAULIN, D
本研究旨在探索一种提高哺乳动物细胞系制备效率的新方法。细胞永生化是通过核内微注射重组DNA构建体实现的，重组DNA构建体由控制编码永生化蛋白基因的组成启动子组成；编码大T抗原和小T抗原的序列在vimentin基因调控元件的下游融合，vimentin基因的激活是绝大多数体外生长细胞的特征。数据显示，使用SV40早期基因的永生化程序的效率可以通过来自人类vimentin (HuVim) 5'序列的控制元件来提高，该序列含有来自CAP位点的核苷酸-878至+93。该重组HuVim 830-T/t用于从多种不同来源的原代培养中创建细胞系：兔、猪和人内皮细胞，兔和牛上皮细胞。我们获得了一组大的t表达细胞，这些细胞保留了分化细胞的特征：结合欧洲巨藻凝集素并合成人内皮细胞的因子VIII；牛输卵管细胞和兔乳腺细胞的细胞角蛋白网络。
The aim of this study was to investigate a new method to enhance the efficiency to create mammalian cell lines. Cell immortalization was achieved by intranuclear microinjection of a recombinant DNA construct composed of a constitutive promoter controlling the genes encoding immortalizing proteins; the sequences coding for the large T and small t antigens were fused down-stream of regulatory elements from the vimentin gene, the activation of which characterizes the vast majority of cells growing in vitro. Data show that the efficiency of the immortalizing procedures using the SV40 early genes could be enhanced by the control elements derived from the human vimentin (HuVim) 5' sequences that contained nucleotides -878 to +93 from the CAP site. This HuVim 830-T/t recombinant was used to create cell lines from numerous primary cultures of different origins: rabbit, porcine and human endothelial cells, rabbit and bovine epithelial cells. A set of large T-expressing cells was derived, and these cells retained characteristics of differentiated cells: binding of Ulex europaeus lectin and synthesis of Factor VIII for human endothelial cells; network of cytokeratin for bovine oviductal cells and rabbit mammary cells.