Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan.
Cytotechnology. 2000 Jul;33(1-3):131-7. doi: 10.1023/A:1008181600197.
The use of the gadd153promoter to induce expression of a reporter geneunder heat stress conditions was investigated,since the results of previous studies have suggestedthat the gadd153promoter is likely to be activated by the indirecteffects of hyperthermia, that is, by DNA damage thatoccurs when reactive oxygen species are produced byheat stress. The optimum temperature for a significantinduction was found to be between 41 and 43 degrees C andincreased expression of the reporter gene was observedat about 24 h after the heat treatment. Under theseconditions, the cell integrity was not alteredmorphologically and the growth stopped temporarily,while the viability was maintained. A second increasein expression occurred at a later stage when the cellswere severely damaged at 43-45 degrees C. Atthese temperatures, the cellular morphology showedsignificant alteration and the growth was stronglyarrested. This is likely to be due to a differentmechanism which could involve DNA repair processes. Itis expected that this method of induction can beexploited to drive the production of a protein ofinterest in a cancer treatment program that includes hyperthermia.
研究了使用 gadd153 启动子在热应激条件下诱导报告基因表达的情况，因为之前的研究结果表明 gadd153 启动子可能是通过热应激产生的活性氧的间接作用而被激活的，即 DNA 损伤。发现显著诱导的最佳温度在 41 到 43 摄氏度之间，并且在热处理后约 24 小时观察到报告基因的表达增加。在这些条件下，细胞完整性在形态上没有改变，生长暂时停止，而活力得以维持。当细胞在 43-45 摄氏度时受到严重损伤时，会出现第二次表达增加。在这些温度下，细胞形态显示出明显的改变，生长受到强烈抑制。这可能是由于涉及 DNA 修复过程的不同机制所致。预计这种诱导方法可用于在包括热疗的癌症治疗计划中驱动感兴趣的蛋白质的产生。
