Quantitative analysis of gene function in the Drosophila embryo.
Quantitative analysis of gene function in the Drosophila embryo.
复制标题
果蝇胚胎中基因功能的定量分析。
DOI：
10.1093/genetics/154.1.273
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发表时间：
2000
期刊：
影响因子：
3.3
通讯作者：
Gergen,JP
中科院分区：
文献类型：
--
作者：
TraceyJr,WD;Ning,X;Klingler,M;Kramer,SG;Gergen,JP
基因产物的特定功能通常取决于它们表达的发育环境。因此，基因功能的研究将受益于系统，允许在模型系统内的基因表达的操作，其中发育的背景是很好地定义。在这里，我们描述了一个系统，允许在正常生理条件下，在早期果蝇胚胎的任何感兴趣的基因的遗传控制过表达。通过使用表达酵母转录因子GAL 4的母体mRNA的果蝇系来实现这种受调节的表达。来自表达GAL4的雌性的胚胎在胚胎发生的胚盘阶段以均匀的水平激活GAL4依赖性UAS转基因。表达水平可以通过使用具有不同水平的母体GAL4活性的品系来定量操纵。特定的表型是通过表达一些不同的发育调节因子而产生的，包括在果蝇胚胎发生过程中通常不起作用的基因。对runt过度表达的反应分析提供了证据，表明这种成对规则分割基因在抑制片段极性基因的转录中具有直接作用。母体GAL4系统将应用于反向遗传实验中基因活性的测量以及对体内基因功能具有定量影响的遗传因素的鉴定。
The specific functions of gene products frequently depend on the developmental context in which they are expressed. Thus, studies on gene function will benefit from systems that allow for manipulation of gene expression within model systems where the developmental context is well defined. Here we describe a system that allows for genetically controlled overexpression of any gene of interest under normal physiological conditions in the early Drosophila embryo. This regulated expression is achieved through the use of Drosophila lines that express a maternal mRNA for the yeast transcription factor GAL4. Embryos derived from females that express GAL4 maternally activate GAL4-dependent UAS transgenes at uniform levels throughout the embryo during the blastoderm stage of embryogenesis. The expression levels can be quantitatively manipulated through the use of lines that have different levels of maternal GAL4 activity. Specific phenotypes are produced by expression of a number of different developmental regulators with this system, including genes that normally do not function during Drosophila embryogenesis. Analysis of the response to overexpression of runt provides evidence that this pair-rule segmentation gene has a direct role in repressing transcription of the segment-polarity gene engrailed. The maternal GAL4 system will have applications both for the measurement of gene activity in reverse genetic experiments as well as for the identification of genetic factors that have quantitative effects on gene function in vivo.