THE RESPONSIVENESS OF A TETRACYCLINE-SENSITIVE EXPRESSION SYSTEM DIFFERS IN DIFFERENT CELL-LINES
THE RESPONSIVENESS OF A TETRACYCLINE-SENSITIVE EXPRESSION SYSTEM DIFFERS IN DIFFERENT CELL-LINES
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DOI：
10.1074/jbc.270.23.14168
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发表时间：
1995-06-09
影响因子：
4.8
通讯作者：
SCHMAUSS, C
中科院分区：
文献类型：
--
作者：
HOWE, JR;SKRYABIN, BV;SCHMAUSS, C
利用四环素敏感诱导表达系统调节两种哺乳动物细胞系神经递质受体基因的表达。多巴胺d -3受体在GH3细胞中稳定表达，GluR6（谷氨酸受体亚基）在人胚胎肾（HEK 293）细胞中稳定表达。他们发现了三个显著的差异。1)在失活状态下，GH3细胞中几乎没有发现d -3受体的表达，而在HER 293细胞中发现了大量GluR6的表达。2)四环素对GH3细胞表达的诱导作用较强，而对HEK 293细胞表达的诱导作用较弱。3)在各克隆细胞系中，共转染的杂交反激活子的表达明显受到四环素响应的调控，在诱导状态下，其mRNA水平在GH3细胞中非常低，在HEK 293细胞中非常高。结果表明，与GH3细胞相比，HEK 293细胞不提供一个可以以四环素反应方式严格控制外源基因表达的细胞环境。
A tetracycline-sensitive inducible expression system was used to regulate the expression of neurotransmitter receptor genes in two mammalian cell lines. The dopamine D-3-receptor was stably expressed in GH3 cells, and GluR6 (a glutamate receptor subunit) was stably expressed in human embryonic kidney (HEK 293) cells. Three striking differences were found. 1) In the inactive state, virtually no D-3-receptor expression was found in GH3 cells, whereas substantial levels of GluR6 expression were found in HER 293 cells. 2) The induction of expression obtained upon removal of tetracycline was robust in GH3 cells but only modest in HEK 293 cells. 3) Whereas in each clonal cell line, the expression of a co-transfected hybrid transactivator is clearly regulated in a tetracycline-responsive manner, in the induced state, its mRNA levels were found to be very low in GH3 cells and very high in HEK 293 cells. The results indicate that, in contrast to GH3 cells, HEK 293 cells do not provide a cellular environment in which the expression of a heterologous gene can be tightly controlled in a tetracycline-responsive manner.