Semiconservative replication

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Semiconservative replication describes the method by which DNA is replicated in all known cells. This method of replication is one of three proposed models[1] [2] of DNA replication:

  • Semiconservative replication would produce two copies that each contained one of the original strands and one new strand.
  • Conservative replication would leave the two original template DNA strands together in a double helix and would produce a copy composed of two new strands containing all of the new DNA base pairs.
  • Dispersive replication would produce two copies of the DNA, both containing distinct regions of DNA composed of either both original strands or both new strands.

The deciphering of the structure of DNA by Watson and Crick in 1953 suggested that each strand of the double helix would serve as a template for synthesis of a new strand. However, there was no way of knowing how the newly synthesized strands might combine with the template strands to form two double helical DNA molecules. The semiconservative model seemed most reasonable since it would allow each daughter strand to remain associated with its template strand. The semiconservative model was confirmed by the Meselson-Stahl experiment and other even more revealing experiments that allowed for autoradiographic visualization of the distribution of old and new strands within replicated chromosomes.

Contents

Testing the semi-conservative theory

Biophysical evidence

The semi-conservative theory can be confirmed by making use of the fact that DNA is made up of nitrogen bases. Nitrogen has an isotope N15 (N14 is the most common isotope) called heavy nitrogen. The experiment that confirms the predictions of the semi-conservative theory makes use of this isotope and runs as follows:

According to the semi-conservative theory, after one replication of DNA, we should obtain 2 hybrid (part N14 part N15) molecules from each original strand of DNA. This would appear as a single line in the test tube. This result would be the same for the dispersive theory. On the other hand, according to the conservative theory, we should obtain one original DNA strand and a completely new one i.e. two fine lines in the test tube placed separately one from the other. Up to this point, either the semi-conservative or the dispersive theories could be truthful, as experimental evidence confirmed that only one line appeared after one replication. In order to conclude between those two, DNA had to be left to replicate again, still in a media containing N14.

In the dispersive theory, after 2 divisions we should obtain a single line, but further up in the test tube, as the DNA molecules become less dense as N14 becomes more abundant in the molecule According to the semi-conservative theory, 2 hybrid molecules and 2 fully N14 molecules should be produced, so two fine lines at different heights in the test tubes should be observed. Experimental evidence confirmed that two lines were observed therefore offering compelling evidence for the semi-conservative theory.

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