| DNA与蛋白质结合的荧光测定 |
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| 引用本文： | 王志珍.DNA与蛋白质结合的荧光测定[J].中国生物化学与分子生物学报,1994,10(4):427-432. |
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| 作者姓名： | 王志珍 |
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| 作者单位： | 中国科学院生物物理所 |
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| 摘 要： | 构建了插入λ阻抑蛋白（Ｒｅｐ）的操纵基因（ＯＲ）和ＢｇｌⅡ识别位点的ＰＢＲ３２２重组质粒。阻抑蛋白与该质粒的相互作用可用ＢｇｌⅡ对它的水解作用引起的ＥＢ荧光变化来研究。在Ｅ．ｃｏｌｉ中表达的Ｒｅｐ表现了与该重组质粒结合的活力。核苷酸序列具精确二重对称性的ＯＲ（ＯＲｃｏｎｓ）对Ｒｅｐ的亲和力比天然的ＯＲ１小。
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| 关 键 词： | DNA－蛋白质结合 λ阻抑蛋白与操纵基因结合 荧光测定 |
| 收稿时间： | 1994-08-20 |
Fluorescence Assay for Binding of DNA to Protein |
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| Wang,Zhi-zhen.Fluorescence Assay for Binding of DNA to Protein[J].Chinese Journal of Biochemistry and Molecular Biology,1994,10(4):427-432. |
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| Authors: | Wang Zhi-zhen |
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| Institution: | (Deparrment of Biochemistry,University of Alberra, Edmonton, Canada |
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| Abstract: | A recombinant PBR322 plasmid with inserts of operator sequence for lambda repressor and Bgl Ⅱ site was constructed. The interaction of lambda repressor to its operator within the recombinant plasmid was evaluated by fluorescence changes of intercalated ethidium bromide produced by Bgl Ⅱ digestion. The crude preperation of lambda repressor expressed in E. Coli showed a significant binding activity to its operator. According to the fluorescence changes, the consensus operator with an exact twofold symmatery sequence showed lower affinity to Rep than natural OR1. |
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| Keywords: | Lambda repressor-operator binding Protein-DNA binding Fluorescence assay |
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