Induction of human high Km 5′-nucleotidase in cultured 293 cells

被引：31

作者：

Gazziola, C ^{[1
]}

Moras, M ^{[1
]}

Ferraro, P ^{[1
]}

Gallinaro, L ^{[1
]}

Verin, R ^{[1
]}

Rampazzo, C ^{[1
]}

Reichard, P ^{[1
]}

Bianchi, V ^{[1
]}

机构：

[1] Univ Padua, Dept Biol, I-35131 Padua, Italy

来源：

EXPERIMENTAL CELL RESEARCH | 1999年 / 253卷 / 02期

关键词：

D O I：

10.1006/excr.1999.4681

中图分类号：

R73 [肿瘤学];

学科分类号：

100214 ;

摘要：

Human 293 cells were stably transfected with a plasmid introducing a receptor for the ecdysone analog muristerone. The cells were further stably transfected with muristerone-inducible expression vectors carrying either the cDNA far the human high K-M 5'-nucleotidase or the coding sequence of the nucleotidase linked to the 5'-end of the sequence for the green fluorescent protein. Upon induction, both types of transfectants overproduced nucleotidase activity in a time- and dose-dependent manner. Western blots gave values close to the expected subunit molecular masses of 65 and 92 kDa, respectively, excluding processing of the induced proteins, Cells induced to overexpress the nucleotidase showed a decreased growth rate and contained smaller pools of each of the four common ribonucleoside triphosphates. They showed no increased resistance to the toxicity of 2-chlorodeoxyadenosine. (C) 1999 Academic Press.

引用

页码：474 / 482

页数：9

共 26 条

[1] Bovine cytosolic IMP/GMP-specific 5'-nucleotidase: cloning and expression of active enzyme in Escherichia coli [J].