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首页> 外文期刊>American Journal of Physiology >The murine SP-C promoter directs type II cell-specific expression in transgenic mice.
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The murine SP-C promoter directs type II cell-specific expression in transgenic mice.

机译：鼠SP-C启动子指导II型细胞在转基因小鼠中的特异性表达。

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摘要

Genomic DNA from the mouse pulmonary surfactant protein C (SP-C) gene was analyzed in transgenic mice to identify DNA essential for alveolar type II cell-specific expression. SP-C promoter constructs extending either 13 or 4.8 kb upstream of the transcription start site directed lung-specific expression of the bacterial chloramphenicol acetyl transferase (CAT) reporter gene. In situ hybridization analysis demonstrated alveolar cell-specific expression in the lungs of adult transgenic mice, and the pattern of 4.8 SP-C-CAT expression during development paralleled that of the endogenous SP-C gene. With the use of deletion constructs, lung-specific, low-level CAT activity was detected in tissue assays of SP-C-CAT transgenic mice retaining 318 bp of the promoter. In transient and stable cell transfection experiments, the 4.8-kb SP-C promoter was 90-fold more active as a stably integrated gene. These findings indicate that 1) the 4.8-kb SP-C promoter is sufficient to direct cell-specific and developmental expression, 2) an enhancer essential for lung-specific expression maps to the proximal 318-bp promoter, and 3) the activity of the 4.8-kb SP-C promoter construct is highly dependent on its chromatin environment.
机译：在转基因小鼠中分析了小鼠肺表面活性蛋白C（SP-C）基因的基因组DNA，以鉴定II型肺泡细胞特异性表达所必需的DNA。在转录起始位点上游延伸13或4.8 kb的SP-C启动子构建体指导细菌氯霉素乙酰转移酶（CAT）报告基因的肺特异性表达。原位杂交分析表明成年转基因小鼠肺中肺泡细胞特异性表达，并且在发育过程中4.8 SP-C-CAT表达的模式与内源SP-C基因的表达模式平行。通过使用缺失构建体，在保留318 bp启动子的SP-C-CAT转基因小鼠的组织检测中检测到了肺特异性，低水平的CAT活性。在瞬时和稳定的细胞转染实验中，4.8-kb SP-C启动子作为稳定整合的基因的活性高90倍。这些发现表明1）4.8-kb SP-C启动子足以指导细胞特异性和发育性表达，2）肺特异性表达所必需的增强子定位到近端318-bp启动子，3） 4.8kb SP-C启动子构建体高度依赖于其染色质环境。

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