Russo V E, Stahl M M, Stahl F W
Proc Natl Acad Sci U S A. 1970 Feb;65(2):363-7. doi: 10.1073/pnas.65.2.363.
The Watson-Crick model for DNA duplex duplication proposes that the two parental chains separate and that each directs the synthesis of a complementary chain with which it is found associated after the duplication act. Previous experiments have left unchallenged alternative models which propose that in any single act of duplication only one of the two parental chains provides information for the synthesis of both new chains. The models are operationally ditinguishable since the former demands that heteroduplexes are destroyed by duplication while the latter anticipates their survivial. We have shown for phage lambda that duplication destroys heterozygotes as predicted by the Watson-Crick model.A stock of lambda containing a high frequency of heterozygotes at the cI locus was prepared by conducting a cross under conditions of depressed DNA synthesis. Particles in this lysate were permitted to duplicate a few times by adsorbing them to a lambda lysogen in a (15)N (13)C medium along with a heteroimmune lambda strain. Emerging lambda particles were separated according to density. The population of particles carrying DNA of parental density retained the initial high heterozygote frequency. Among particles which had duplicated, 80 per cent or more of the heterozygotes had disappeared.
DNA双链复制的沃森-克里克模型提出,两条亲代链分离,且每条链都指导合成一条互补链,复制完成后与之结合在一起。之前的实验并未对其他模型提出质疑,这些模型认为在任何一次单独的复制过程中,两条亲代链中只有一条为两条新链的合成提供信息。这些模型在操作上是可区分的,因为前者要求杂合双链在复制过程中被破坏,而后者预期它们会存活下来。我们已经证明,对于噬菌体λ来说,复制会如沃森-克里克模型所预测的那样破坏杂合子。通过在DNA合成受抑制的条件下进行杂交,制备了一种在cI位点具有高频杂合子的λ噬菌体原种。通过将该裂解物中的颗粒与一种异源免疫的λ菌株一起吸附到处于(15)N(13)C培养基中的λ溶原菌上,使这些颗粒进行几次复制。根据密度分离出新生的λ颗粒。携带亲代密度DNA的颗粒群体保持了最初的高杂合子频率。在已经复制的颗粒中,80%或更多的杂合子消失了。