Harshey R M, McKay R, Bukhari A I
Cell. 1982 Jun;29(2):561-71. doi: 10.1016/0092-8674(82)90172-6.
Transposable genetic elements can insert into DNA sites that have no homology to themselves. Evidence that there is a physical linkage between a transposable element and its target DNA sequence during transposition comes from studies on bacteriophage Mu DNA transposition in which plasmids containing Mu DNA have been shown to attach to host DNA. We report the isolation of key structures, seen after induction of Mu DNA replication, after cloning lac operator into Mu DNA and using the lac repressor-operator interaction to trap Mu DNA on nitrocellulose filters. We have localized Mu sequences within these structures in the electron microscope by visualizing the lac operator-repressor interaction after binding with ferritin-conjugated antibody. This analysis shows that key structures contain replicating Mu DNA linked to non-Mu DNA and that replication can begin at either end of Mu.
可转座遗传元件能够插入与其自身没有同源性的DNA位点。转座过程中转座元件与其靶DNA序列之间存在物理连接的证据来自对噬菌体Mu DNA转座的研究,其中含有Mu DNA的质粒已被证明可附着于宿主DNA。我们报告了在诱导Mu DNA复制后、将lac操纵基因克隆到Mu DNA中并利用lac阻遏物-操纵基因相互作用将Mu DNA捕获在硝酸纤维素滤膜上之后所观察到的关键结构的分离。我们通过用铁蛋白偶联抗体结合后观察lac操纵基因-阻遏物相互作用,在电子显微镜下将Mu序列定位在这些结构内。该分析表明,关键结构包含与非Mu DNA相连的正在复制的Mu DNA,并且复制可以从Mu的任一端开始。