Holt J T
Department of Cell Biology, Vanderbilt University, Nashville, Tennessee 37232.
Ann N Y Acad Sci. 1992 Oct 28;660:88-94. doi: 10.1111/j.1749-6632.1992.tb21061.x.
We have employed antisense methods to study the transcriptional functions of c-fos protein (Fos). Clones expressing inducible anti-fos RNA have been employed to inhibit c-fos expression, resulting in activation of c-fos transcription by inhibiting its normal repressor function. The sites of negative regulation by Fos have been mapped using this antisense mapping method which demonstrates that the serum response element represents the major site of repression by endogenous c-fos protein. A similar strategy (antisense cloning) has been employed to clone four target genes that are Fos dependent. These cDNAs encode mRNAs that are rapidly induced by serum (although this induction is blocked by cycloheximide) but are blocked by induction of anti-fos RNA. These inhibitory methods of studying transcription factor function are extremely useful for transcription factors (like Fos) that require cooperation with other factors to modulate gene transcription.
我们采用反义方法来研究c-fos蛋白（Fos）的转录功能。已使用表达可诱导性抗Fos RNA的克隆来抑制c-fos表达，通过抑制其正常的阻遏功能导致c-fos转录激活。利用这种反义定位方法已确定了Fos负调控的位点，该方法表明血清反应元件是内源性c-fos蛋白的主要抑制位点。已采用类似策略（反义克隆）来克隆四个Fos依赖性靶基因。这些cDNA编码的mRNA可被血清快速诱导（尽管这种诱导被放线菌酮阻断），但被抗Fos RNA的诱导所阻断。这些研究转录因子功能 的抑制方法对于需要与其他因子协同调节基因转录的转录因子（如Fos）极为有用。
