Semiconservative replication
- 1. Semiconservative replication ⚫ It is crucial that the genetic material is reproduced accurately. When Watson and Crick worked out the double-helix structure of DNA in 1953, they recognized that the complementary nature of the two strands-A paired with T and G paired with C-might play an important role in its replication. Because the two polynucleotide strands are joined only Genetics by hydrogen bonds, they are able to separate without requiring breakage of covalent bonds. ⚫ If the two strands of a parental double helix of DNA are separated, the base sequence of each parental strand could serve as a template for the synthesis of a new complementary strand, producing two identical progeny double helices. This process is called semiconservative replication because the parental double helix is half conserved, each parental single strand remaining intact. ⚫ The alternative methods are conservative and dispersive. ⚫ In conservative replication, the whole original double helix acts as a template for a new one, one daughter molecule would consist of the original parental DNA, and the other daughter would be totally new DNA. In dispersive replication, some parts of the original double helix are conserved, and some parts are not. In this model, the parental double helix is broken into double-stranded DNA segments and just like conservative mode of replication acts as templates for the synthesis of new double- stranded DNA segments. The segments then reassemble into complete
- 2. DNA double helices, each with parental and progeny DNA segments interspersed. *The Meselson-Stahl experiment showed that DNA replicates by a semiconservative mechanism In this experiment, E. coli cells initially were grown in a medium containing ammonium salts prepared with heavy nitrogen (15N) until the entire cellular DNA was labeled. After the cells were transferred to a medium containing the normal light isotope (14N), samples were removed periodically from the cultures and the DNA in each sample was analyzed by equilibrium density-gradient centrifugation. This technique can separate heavy-heavy (15N-15N), light-light (14N-14N), and heavy-light (15N-14N) duplexes into distinct bands.
- 3. After one generation of growth, the entire extracted DNA had the density of 15N-14N DNA. After two generations, approximately half the DNA had the density of 15N- 14N DNA; the other half had the density of 15N-14N DNA. With additional generations, a large fraction of the extracted DNA consisted of 14N-14N duplexes; 15N-15N duplexes never appeared. These results match the predicted pattern for the semi-conservative replication mechanism. Meselson and Stahl experimentally demonstrated the semi-conservative replication of DNA in E. coli in 1958. If the parental DNA carries a heavy density label because the organism has been grown in medium containing a suitable isotope (such as 15N), its strands can be distinguished from those that are synthesized when the organism is transferred to a medium containing normal light isotopes.