利用鸡红细胞核提取物说明dna -蛋白相互作用的实验

Beatriz Garat
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引用次数: 2

摘要

电泳迁移率转移试验是一种简单、快速的观察dna -蛋白特异性相互作用存在的方法,是一种有用的教学实验。在这里描述的实验中,学生准备核提取物,DNA探针,并可以成功地产生特定的DNA-蛋白质复合物,并估计表征相互作用的物理化学参数。所描述的系统易于执行。合成所需的寡核苷酸和标记的脱氧核糖核苷酸用于标记探针是实验的主要成本,但由于合成的寡核苷酸可以用于几个课程,标记探针所需的标记的脱氧核糖核苷酸的数量很少,因此在一个班级进行实验相对便宜。
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An experiment illustrating DNA-protein interactions using nuclear extracts from chicken erythrocytes

The electrophoretic mobility shift assay is a simple and rapid method for visualizing the existence of specific DNA-protein interactions and is a useful teaching experiment. In the experiment described here the students prepare nuclear extracts, a DNA probe, and can successfully produce specific DNA-protein complexes and estimate physicochemical parameters characterizing the interaction. The system described is easy to perform. The synthesis of the oligonucleotides required and the labeled deoxyribonucleotide used for labeling the probe are the main costs of the experiment, but since the synthesized oligonucleotides can be used in several courses and the amount of labeled deoxyribonucleotide required for labeling the probe is small the experiment is relatively inexpensive to put on with a class.

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