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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >An efficient promoter trap for detection of patterned gene expression and subsequent functional analysis in Drosophila
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An efficient promoter trap for detection of patterned gene expression and subsequent functional analysis in Drosophila

机译：一个有效的启动子陷阱，用于检测果蝇中模式化的基因表达并进行后续功能分析

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摘要

Transposable elements have been used in Drosophila to detect gene expression, inactivate gene function, and induce ectopic expression or overexpression. We have combined all of these features in a single construct. A promoterless GAL4 cDNA is expressed when the construct inserts within a transcriptional unit, and GAL4 activates a GFP-encoding gene present in the same transposon. In a primary screen, patterned gene expression is detected as GFR fluorescence in the live progeny of dysgenic males. Many animals expressing GFP in distinct patterns can be recovered with relatively little effort. As expected, many insertions cause loss of function. After insertion at a genomic location, specific parts of the transposon can be excised by FLP recombinase, thus allowing it to induce conditional misexpression of the tagged gene. Therefore, both gain- and loss-of-function studies can be carried out with a single insertion in a gene identified by virtue of its expression pattern. Using this promoter trap approach, we have identified a group of cells that innervate the calyx of the mushroom body and could thus define a previously unrecognized memory circuit.
机译：果蝇中已将转座元件用于检测基因表达，失活基因功能并诱导异位表达或过表达。我们将所有这些功能组合在一个结构中。当构建体插入转录单位内时，表达无启动子的GAL4 cDNA，而GAL4激活同一转座子中存在的GFP编码基因。在初步筛选中，检测到图案化的基因表达为雄性不育男性的活后代中的GFR荧光。以不同模式表达GFP的许多动物只需相对较少的努力即可恢复。如预期的那样，许多插入会导致功能丧失。在基因组位置插入后，转座子的特定部分可以通过FLP重组酶切除，从而使其能够诱导标记基因的条件错误表达。因此，功能获得和功能丧失研究都可以在通过基因表达方式鉴定的基因中插入一个基因而进行。使用这种启动子陷阱方法，我们已经鉴定出了一组能使蘑菇体的萼神经支配的细胞，从而可以定义一个以前无法识别的记忆电路。

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