HIGHLY INDUCIBLE SYNTHESIS OF HETEROLOGOUS PROTEINS IN EPITHELIAL-CELLS CARRYING A GLUCOCORTICOID-RESPONSIVE VECTOR
HIGHLY INDUCIBLE SYNTHESIS OF HETEROLOGOUS PROTEINS IN EPITHELIAL-CELLS CARRYING A GLUCOCORTICOID-RESPONSIVE VECTOR
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DOI：
10.1016/0378-1119(92)90687-k
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发表时间：
1992-02-15
期刊：
影响因子：
3.5
通讯作者：
FASEL, N
中科院分区：
文献类型：
--
作者：
HIRT, RP;POULAINGODEFROY, O;FASEL, N
描述了一种糖皮质激素响应性载体，其允许互补DNA（cDNA）在稳定转染的哺乳动物细胞系中的高度可诱导表达。该载体pLK-neo由变体小鼠乳腺肿瘤病毒长末端重复启动子组成，含有激素调节元件、在猿猴病毒40转录单位中的遗传霉素抗性编码基因和异源cDNA的多接头插入位点，该载体用于表达多聚免疫球蛋白（poly-Ig）受体和胸腺细胞标记物Thy-1，在Madin-Darby犬肾（MDCK）细胞和鼠成纤维细胞L细胞中。在MDCK细胞中观察到高水平的聚Ig受体或Thy-1 mRNA积累，以响应地塞米松，同时蛋白质合成增加10至200倍，这取决于重组蛋白和转染的细胞克隆。
A glucocorticoid-responsive vector is described which allows for the highly inducible expression of complementary DNAs (cDNAs) in stably transfected mammalian cell lines. This vector, pLK-neo, composed of a variant mouse mammary tumor virus long terminal repeat promoter, containing a hormone regulatory element, a Geneticin resistance-encoding gene in a simian virus 40 transcription unit, and a polylinker insertion site for heterologous cDNAs, was used to express the polymeric immunoglobulin (poly-Ig) receptor and the thymocyte marker, Thy-1, in Madin-Darby canine kidney (MDCK) cells and in murine fibroblast L cells. A high level of poly-Ig receptor or Thy-1 mRNA accumulation was observed in MDCK cells in response to dexamethasone with a parallel ten- to 200-fold increase in protein synthesis depending on the recombinant protein and the transfected cell clone.