Raman V, Rand K N, Hill R J
CSIRO Division of Biomolecular Engineering, North Ryde, NSW, Australia.
Biochim Biophys Acta. 1994 Oct 18;1219(2):441-8. doi: 10.1016/0167-4781(94)90070-1.
Regulatory elements from the arabinose operon of Escherichia coli were transfected into Drosophila melanogaster Schneider line 2 cells to test their ability to function in animal cells. A construct containing an araC fusion gene (encoding AraC protein) under the control of an act5C (encoding actin) promoter and a construct containing a lacZ fusion gene (reporter gene) also under the control of an act5C promoter were used to build a regulatory circuit in the D. melanogaster cells. We have demonstrated that a AraC fusion protein can be synthesised in Schneider cells where it is able to repress the transcription of the reporter gene containing AraC-binding sites inserted between the transcription start point and the initiation codon. The reporter gene activity can be further modulated by the addition of arabinose to the medium.
将来自大肠杆菌阿拉伯糖操纵子的调控元件转染到黑腹果蝇施耐德2号线细胞中，以测试它们在动物细胞中的功能能力。使用一个在act5C（编码肌动蛋白）启动子控制下包含araC融合基因（编码AraC蛋白）的构建体和一个同样在act5C启动子控制下包含lacZ融合基因（报告基因）的构建体，在黑腹果蝇细胞中构建一个调控回路。我们已经证明，在施耐德细胞中可以合成AraC融合蛋白，它能够抑制包含插入转录起始点和起始密码子之间的AraC结合位点的报告基因的转录。通过向培养基中添加阿拉伯糖，可以进一步调节报告基因的活性。
