Robinett C C, Dunaway M
Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA.
Methods. 1999 Feb;17(2):151-60. doi: 10.1006/meth.1998.0726.
Transcriptional regulation is a complex process that requires cooperation between specific DNA sequence elements, the DNA-binding proteins that bind to these sequences, the general transcriptional machinery, and chromatin. Oocyte microinjection offers a technique to study the integrated transcription process while still providing the opportunity to experimentally perturb this process. We describe here techniques for manipulating DNA templates and the protein complement of the oocyte to study multiple facets of transcriptional regulation. We present sample results showing that the GAL4-VP16 fusion activator is sensitive to distance in constructs containing only a minimal promoter, but can activate transcription at greater distances when proximal promoter elements are present.
转录调控是一个复杂的过程，需要特定DNA序列元件、与这些序列结合的DNA结合蛋白、通用转录机制和染色质之间的协同作用。卵母细胞显微注射提供了一种研究整合转录过程的技术，同时仍提供了通过实验干扰这一过程的机会。我们在此描述了操纵DNA模板和卵母细胞蛋白质成分以研究转录调控多个方面的技术。我们展示的样本结果表明，GAL4-VP16融合激活剂在仅含有最小启动子的构建体中对距离敏感，但当存在近端启动子元件时，它可以在更远的距离激活转录。
