Regulation of mouse ornithine decarboxylase gene expression in a macrophage-like cell line: synergistic induction by bacterial lipopolysaccharide and cAMP.
Regulation of mouse ornithine decarboxylase gene expression in a macrophage-like cell line: synergistic induction by bacterial lipopolysaccharide and cAMP.
复制标题
巨噬细胞样细胞系中小鼠鸟氨酸脱羧酶基因表达的调节：细菌脂多糖和 cAMP 的协同诱导。
DOI：
10.1016/s0006-291x(05)81198-2
复制
发表时间：
1991
影响因子：
3.1
通讯作者：
Taffet,SM
中科院分区：
文献类型：
--
作者：
Zheng,SA;McElwain,CM;Taffet,SM
研究了小鼠鸟氨酸脱羧酶基因的启动子/增强子区域对各种刺激的响应能力。将该区域亚克隆成多个片段，并将这些片段插入表达载体pBLCAT 3上氯霉素乙酰转移酶基因的前面。将这些ODC/CAT构建体转染到小鼠巨噬细胞样细胞系RAW 264中。用细菌脂多糖、8-溴cAMP或两者刺激转染的细胞，然后分析氯霉素乙酰转移酶活性。当在脂多糖和8-溴cAMP组合处理的细胞中测试-90至+12（相对于转录起始位点）的序列时，获得了最佳的诱导型氯霉素乙酰转移酶表达。一个假定的环AMP反应元件位于-48改变了定点诱变，但这些改变并没有减少活性的刺激与脂多糖和8-溴环腺苷酸。
The ability of the promotor/enhancer region of the mouse ornithine decarboxylase gene to respond to various stimuli was studied. This region was subcloned into multiple fragments and these were inserted in front of the chloramphenicol acetyltransferase gene on an expression vector, pBLCAT3. These ODC/CAT constructs were transfected into a mouse macrophage-like cell line, RAW264. The transfected cells were stimulated by bacterial lipopolysaccharide, 8-bromo cAMP or both followed by analysis of chloramphenicol acetyltransferase activity. Optimal inducible chloramphenicol acetyltransferase expression was obtained when sequences from −90 to + 12 (with respect to the transcriptional start site) were tested in cells treated with a combination of lipopolysaccharide and 8-bromo cAMP. A putative cyclic AMP response element located at -48 was altered by site-directed mutagenesis but these alterations did not diminish activity in response to stimulation with lipopolysaccharide and 8-bromo CAMP.