大肠杆菌lacZ基因融合在哺乳动物细胞中的表达与调控。

C V Hall, P E Jacob, G M Ringold, F Lee
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引用次数: 0

摘要

大肠杆菌lacZ基因与含有猴病毒40早期启动子或小鼠乳腺肿瘤病毒(MMTV)启动子的DNA片段的基因融合指导了Cos 7猴细胞和小鼠ltk细胞中功能性β -半乳糖苷酶的合成。在转染72 h后或在稳定的转化子中测量酶活性。采用敏感的β -半乳糖苷酶测定法测定基因表达并优化dna介导转染的效率。当lacZ与激素调节的MMTV启动子融合时,糖皮质激素刺激β -半乳糖苷酶的产生。
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Expression and regulation of Escherichia coli lacZ gene fusions in mammalian cells.

Gene fusions between the Escherichia coli lacZ gene and DNA segments containing the simian virus 40 early promoter or the mouse mammary tumor virus (MMTV) promoter direct the synthesis of functional beta-galactosidase in Cos 7 monkey cells and mouse Ltk-cells. Enzymatic activity was measured either 72 h after transfection or in stable transformants. The sensitive beta-galactosidase assay was used to measure gene expression and to optimize the efficiency of DNA-mediated transfection. Glucocorticoids stimulated the production of beta-galactosidase when lacZ was fused to the hormonally regulated MMTV promoter.

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