Shevelev A Ia, Kondratov R V, Rybalkin I N, Prasolov V S
Mol Biol (Mosk). 1992 Jan-Feb;26(1):208-17.
Expression of urokinase in murine and rat cells was performed by two recombinant constructs, one containing cDNA and the other--hybrid (cDNA/genome) variant of human urokinase gene conserving 7 introns of 10, in the eukaryotic retrovirus vector pPS-3-neo. DNA of both constructs was introduced into packaging cell line psi 2 by a standard Ca-phosphate transfection technique. Infection of mouse and rat fibroblasts BALB/c 3T3 and Rat I with virus particles, produced by transfected psi 2 cells, led to an integration into the host genome of one or two recombinant proviral copies. Stable expression and secretion into the culture medium of glycosylated high molecular weight human urokinase was observed for both cell types. For the hybrid gene construct, precise excision of intervening sequences was shown during transferring of genetic material from packaging to recipient cells.
通过两种重组构建体在小鼠和大鼠细胞中表达尿激酶，一种构建体包含互补DNA（cDNA），另一种是人类尿激酶基因的杂交（cDNA/基因组）变体，保留了10个内含子中的7个，构建于真核逆转录病毒载体pPS-3-neo中。通过标准的磷酸钙转染技术将两种构建体的DNA导入包装细胞系psi 2。用转染的psi 2细胞产生的病毒颗粒感染小鼠和大鼠成纤维细胞BALB/c 3T3和大鼠I型细胞，导致一两个重组前病毒拷贝整合到宿主基因组中。两种细胞类型均观察到糖基化的高分子量人尿激酶在培养基中的稳定表达和分泌。对于杂交基因构建体，在遗传物质从包装细胞转移到受体细胞的过程中，显示出间隔序列的精确切除。
