Roman G, Endo K, Zong L, Davis R L
Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX 77030, USA.
Proc Natl Acad Sci U S A. 2001 Oct 23;98(22):12602-7. doi: 10.1073/pnas.221303998.
We have developed a method for turning on and off the expression of transgenes within Drosophila in both time and space. Two different enhancer detector elements carrying an RU486-inducible form of the yeast transcription factor GAL4 were constructed and used to generate enhancer detector lines. These lines were screened for RU486-inducible reporter gene expression in the adult head. We identified lines that exhibit inducible expression in many cell and tissue types, verifying that the elements respond to nearby enhancers. No expression was detected in the absence of the ligand. The P[Switch1] element responded to genomic enhancers less efficiently than P[Switch2] but produced more specific patterns of expression. Two P[Switch] lines were used to ablate fat body tissue in adult females through the induced expression of diphtheria toxin. These females were sterile, which correlates with fat body loss, and they died prematurely.
我们已经开发出一种方法,可在果蝇体内在时间和空间上开启和关闭转基因的表达。构建了两种携带酵母转录因子GAL4的RU486诱导形式的不同增强子检测元件,并用于生成增强子检测系。对这些品系进行筛选,以检测成年果蝇头部中RU486诱导的报告基因表达。我们鉴定出在许多细胞和组织类型中表现出诱导表达的品系,证实这些元件对附近的增强子有反应。在没有配体的情况下未检测到表达。P[Switch1]元件对基因组增强子的反应效率低于P[Switch2],但产生了更特异的表达模式。使用两个P[Switch]品系通过诱导表达白喉毒素来消融成年雌性果蝇的脂肪体组织。这些雌性果蝇不育,这与脂肪体缺失相关,并且它们过早死亡。