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利用组织纤溶酶原激活物基因表达对转染的哺乳动物细胞进行中性选择。

Neutral selection of transfected mammalian cells using tissue plasminogen activator gene expression.

作者信息

Rogers P, Fisher R, Guyden J

机构信息

Department of Microbiology and Immunology, University of California, Berkeley 94720.

出版信息

Gene. 1989 Sep 1;81(1):151-8. doi: 10.1016/0378-1119(89)90345-4.

Abstract

The gene that produces the proteolytic enzyme, tissue plasminogen activator (tPA), was used as a selectable marker for transfection. Cells that were successfully transfected with plasmids containing a gene for tPA (tpa) produce plaques in the caseinolysis assay. Two tpa-containing plasmids were constructed and used to transfect a variety of cell types. One plasmid contained the promoter region of simian virus 40 and the other contained the Moloney murine leukemia virus promoter. No significant difference in transfection frequencies was found for the two plasmids. However, 80% of the cell types tested produced plaques. This system allows for the identification and isolation of transfected cells under conditions that are not lethal to nontransfected cells.

摘要

产生蛋白水解酶组织型纤溶酶原激活剂(tPA)的基因被用作转染的选择标记。成功用含有tPA基因(tpa)的质粒转染的细胞在酪蛋白溶解试验中会形成噬菌斑。构建了两个含tpa的质粒并用于转染多种细胞类型。一个质粒含有猿猴病毒40的启动子区域,另一个含有莫洛尼鼠白血病病毒启动子。这两个质粒的转染频率没有显著差异。然而,80%的受试细胞类型产生了噬菌斑。该系统能够在对未转染细胞无致死性的条件下鉴定和分离转染细胞。

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