克雷复合酶与DNA复合的结构在特定位点的复合突触中
相关实验视频
Updated: Nov 1, 2026

07:06
Principles of Site-Specific Recombinase (SSR) Technology
Published on: May 29, 2008
F Guo1, D N Gopaul, G D van Duyne
1Johnson Research Foundation and Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, Philadelphia 19104, USA.
Nature
|September 4, 1997
概括
菌体Cre复合酶通过结合loxP位点促进了基因重组. 它的晶体结构揭示了反应中间体,挑战了现有的特定地点重组的现有模型.
科学领域:
- 分子生物学分子生物学
- 结构生物学 结构生物学
- 遗传学 是一个遗传学.
背景情况:
- 局部特异性DNA重组使遗传重组成为病毒整合,切除和染色体分离的关键.
- 重组酶酶通过识别特定的DNA序列并催化链交换来调解这一过程.
研究的目的:
- 为了阐明菌体的结构机制,CRE重组酶介导的局部特异性重组.
- 为了研究在Cre-loxP重组过程中形成的中间体的结构基础.
主要方法:
- 使用X射线晶体学以2.4 Å分辨率确定Cre重组酶与loxP DNA基质结合的结构.
- 结构分析的重点是识别重组反应期间的关键中间体和构造变化.
主要成果:
- 晶体结构揭示了一种反应中间体,其中一个Cre分子与切割的DNA基质形成了共价3'-酸铁链接.
- 观察到一个涉及四个重组酶和两个loxP位点的突触结构,类似于霍莱德 - 交叉路口中间体.
- 观察到的结构挑战了需要显著四级结构变化的模型,这表明微妙的全变化协调了反应.
结论:
- 克雷-洛克斯P复合体结构为我们提供了在分子水平上对特定位点重组的机制的见解.
- 在Cre重组酶中微妙的全变化可能调节DNA分裂和链交换,提供了替代的机械模型.
相关概念视频
Conservative Site-specific Recombination and Phase Variation
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
Homologous Recombination
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
Homologous Recombination
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
The Replisome
DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
The Replisome
DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
Overview of Transposition and Recombination
Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...

