在子细胞中通过大肠杆菌乳液抑制剂对稳定集成的氨基醇乙转移酶基因进行严格调节
J Figge1, C Wright, C J Collins
1Division of Neoplastic Disease Mechanisms, Dana-Farber Cancer Institute, Boston, Massachusetts 02115.
Cell
|March 11, 1988
概括
研究人员创造了具有可诱导基因表达的子细胞. 当这些细胞接触到异基β-D-thiogalactoside (IPTG) 时,它们会激活一个记者基因,从而在灵长类细胞中显示出精确的遗传控制.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 细胞生物学 细胞生物学
背景情况:
- 记者基因稳定整合到灵长类染色体中对于基因表达研究至关重要.
- 拉克抑制器-操作器系统为诱导性基因调节提供了一个潜在的机制.
研究的目的:
- 为研究可诱导基因表达生成子细胞系.
- 为了研究在灵长类细胞中使用lac抑制器-操作器系统进行转录控制的可行性.
主要方法:
- 产生子细胞系与集成的乳酸抑制剂和氨基醇乙转移酶 (CAT) 基因.
- 细胞暴露于异甲基β-D-thiogalactoside (IPTG) 以诱导CAT基因表达.
- 在不同的IPTG度和退出条件下监测CAT活动.
主要成果:
- 细胞在暴露于IPTG时表现出CAT+表型,表明成功诱导基因.
- CAT活动增加了多达60倍,显示出显著的转录控制.
- 诱导细胞在去除IPTG后恢复到CAT-表型,显示可逆性.
结论:
- 拉克抑制器-操作器复合体可以有效地与灵长类动物染色体结合.
- 这个系统在子细胞中提供了严格的转录抑制和强大的诱导.
- 开发出来的细胞系是哺乳动物系统中诱导基因表达研究的宝贵工具.
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