Differential replication of circular DNA molecules co-injected into early Xenopus laevis embryos

Nucleic Acids Res. 1989 Jul 25;17(14):5793-808. doi: 10.1093/nar/17.14.5793.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Blastocyst / metabolism
  • DNA Replication*
  • DNA, Circular / biosynthesis*
  • DNA, Circular / genetics
  • DNA, Circular / isolation & purification
  • Embryo, Nonmammalian / metabolism*
  • Female
  • Kinetics
  • Phosphorus Radioisotopes
  • Templates, Genetic
  • Xenopus laevis

Substances

  • DNA, Circular
  • Phosphorus Radioisotopes