真核生物转录机理与调控详解.ppt

Actually, the activity of the different polymerases was determined using α-amanitin. Alpha-amanitin is produced by a kind of mushroom and is deadly poisonous to human beings. 实际上，不同聚合酶的活性是使用了一种叫做α鹅膏蕈碱的毒素来测定的，这种毒素是一种蘑菇产生的，对人的毒性非常大。 In order to see which genes are transcribed by which polymerase, cells are exposed to α-amanitin. It was found that, under very high concentrations of α-amanitin, only rRNA precursor is produced. This means that rRNA precursor is produced by RNA polymerase I. 为了弄清哪些基因由哪种聚合酶转录，细胞被暴露在α鹅膏蕈碱中。暴露在很高浓度α鹅膏蕈碱的细胞只产生rRNA前体，意味着这正是RNA聚合酶I所转录的基因。 Then, cells expose to low concentrations of the toxin also produce tRNA and other small RNAs like the 5S rRNA, suggesting that these are the genes transcribed RNA polymerase III. mRNA is not produced even at low concentrations of α-amantin, suggesting that RNA polymerase II is responsible for transcription of mRNA. 而暴露在低浓度毒素中的细胞还能产生tRNA和其它小RNA如5S rRNA，意味着这些基因是由RNA聚合酶III转录的。即使在低浓度α鹅膏蕈碱下mRNA也不能产生，说明RNA聚合酶II负责转录mRNA。 The second experiment we are going to look at is the one to find out the modularity of specific transcription factors. Modularity means that a protein is composed of separate modules. For example, a specific transcription factor is composed of one DNA binding module and one transcription activation module. When two such proteins exchange their modules, they will still be able to function. 我们要看的第二个实验研究是关于怎么发现特异转录因子模块化的。模块化的意思是一种蛋白质由不同的模块组成，例如，特异转录因子含有一个DNA结合模块和一个分开的激活模块，如果两种蛋白交换它们的模块，交换后两者仍然具有功能。 Before explaining the experiments to demonstrate the modularity of specific transcription factors, let’s first understand what a reporter gene is. A reporter gene is a gene whose transcriptional and/or translational products can be easily detected and measured. For example, genes coding for GFP and RFP proteins are very frequently used as reporter genes. GFP stands for green fluorescent protein. This protein can emit green fluorescent light under visible light. 在解释特异转录因子的模块化实验研究内容之前，我们先来了解一下什么是报告基因