Marini N J, Hiriyanna K T, Benbow R M
Department of Zoology, Iowa State University, Ames 50011.
Nucleic Acids Res. 1989 Jul 25;17(14):5793-808. doi: 10.1093/nar/17.14.5793.
Replication of co-injected supercoiled DNA molecules in fertilized Xenopus eggs was monitored through the blastula stage of development. The extent of replication, as measured by 32P-dTMP incorporation into form I DNA, was directly proportional to the number of molecules, rather than the size, of the plasmid injected. Although only a small fraction of molecules of either template was replicated, incorporation was predominantly into full length daughter molecules. Over at least a 20-fold concentration range of microinjected DNA, injection of equal masses of DNA resulted in greater incorporation into the smaller form I DNA present in molar excess. The extent of incorporation into supercoiled DNA for a particular plasmid was apparently independent of the concentration of a second, co-injected plasmid. The relative extents of replication of co-injected supercoiled templates could be altered simply by changing the molar ratios of the templates.
通过非洲爪蟾受精卵发育的囊胚期监测共注射的超螺旋DNA分子的复制情况。以32P-dTMP掺入I型DNA来衡量的复制程度,与注射的质粒分子数量直接成正比,而非质粒大小。尽管两种模板中只有一小部分分子被复制,但掺入主要发生在全长子代分子中。在至少20倍浓度范围内的显微注射DNA中,注射等量质量的DNA会导致更多地掺入摩尔过量的较小I型DNA中。特定质粒掺入超螺旋DNA的程度显然与共注射的第二种质粒的浓度无关。共注射的超螺旋模板的相对复制程度可以通过简单地改变模板的摩尔比来改变。