Distinct expression patterns detected within individual tissues by the GAL4 enhancer trap technique
Distinct expression patterns detected within individual tissues by the GAL4 enhancer trap technique
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DOI：
10.1139/g96-023
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发表时间：
1996-02-01
期刊：
影响因子：
3.1
通讯作者：
Boulianne, GL
中科院分区：
文献类型：
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作者：
Gustafson, K;Boulianne, GL
为了鉴定在发育过程中特定细胞类型或组织中表达的基因，我们构建了增强子捕获品系，在这些品系中，酵母转录激活因子GAL4在果蝇基因组中被移动。GAL4品系是一个由GAL4及其靶标——上游激活序列（UAS）组成的两部分系统的一部分。通过将单个GAL4品系与在UAS转录控制下携带报告基因lacZ的果蝇杂交，然后进行组织化学和免疫细胞化学染色，来检测GAL4的表达模式。在此，我们展示了这项筛选的结果以及GAL4品系的特征，这些品系在果蝇发育过程中，包括胚胎发生、卵子发生和成虫盘发育过程中，显示出不同的基因表达模式。然而，我们无法鉴定出在生殖系内或早期胚胎发生过程中表达的GAL4品系。此外，与先前的结果一致，我们发现GAL4增强子捕获技术的转座频率比lacZ增强子捕获筛选所报道的要低得多。综上所述，这些结果既展示了GAL4增强子捕获技术在鉴定发育过程中独特基因表达模式方面的优势，也揭示了其劣势。
To identify genes that are expressed in specific cell types or tissues during development, we generated enhancer-trap lines in which the yeast transcriptional activator, GAL4, was mobilized throughout the Drosophila genome. The GAL4 lines are part of a two-part system involving GAL4 and its target, the upstream activating sequence (UAS). Detection of GAL4 expression patterns was achieved by crossing individual GAL4 lines with flies carrying the reporter gene lacZ under the transcriptional control of the UAS followed by histochemical and immunocytochemical staining. Here, we present the results of this screen and the characterization of GAL4 lines that show distinct patterns of gene expression during Drosophila development, including embryogenesis, oogenesis, and imaginal disc development. However, we were unable to identify GAL4 lines that were expressed within the germ line or during early embryogenesis. Furthermore, consistent with previous results, we found that the GAL4 enhancer trap technique had a much lower frequency of transposition than has been reported for lacZ enhancer trap screens. Taken together, these results demonstrate both the strengths and weaknesses of the GAL4 enhancer trap technique for identifying unique patterns of gene expression during development.