thumb|Overview of experiment and observations|357x357px
The Hershey–Chase experiments, also called the Blender experiment, were a series of experiments conducted in 1952 by Alfred Hershey and Martha Chase that helped to confirm that DNA is genetic material.
thumb|Scientists Martha Chase and Alfred Hershey
While DNA had been known to biologists since 1869, many scientists still assumed at the time that proteins carried the information for inheritance because DNA appeared to be an inert molecule, and, since it is located in the nucleus, its role was considered to be phosphorus storage. In their experiments, Hershey and Chase showed that when bacteriophages, which are composed of DNA and protein, infect bacteria, their DNA enters the host bacterial cell, but most of their protein does not. Hershey and Chase and subsequent discoveries all served to prove that DNA is the hereditary material.
Hershey shared the 1969 Nobel Prize in Physiology or Medicine with Max Delbrück and Salvador Luria for their "discoveries concerning the genetic structure of viruses".
Historical background
In the early twentieth century, biologists thought that proteins carried genetic information. This was based on the belief that proteins were more complex than DNA. Phoebus Levene's influential "tetranucleotide hypothesis", which incorrectly proposed that DNA was a repeating set of identical nucleotides, supported this conclusion. The results of the Avery–MacLeod–McCarty experiment, published in 1944, suggested that DNA was the genetic material, but there was still some hesitation within the general scientific community to accept this, which set the stage for the Hershey–Chase experiment.
Hershey and Chase, along with others who had done related experiments, confirmed that DNA was the biomolecule that carried genetic information. Before that, Oswald Avery, Colin MacLeod, and Maclyn McCarty had shown that DNA led to the transformation of one strain of Streptococcus pneumoniae to another. The results of these experiments provided evidence that DNA was the biomolecule that carried genetic information.
Methods and results
thumb|Structural overview of T2 phage
Hershey and Chase needed to be able to examine different parts of the phages they were studying separately, so they needed to distinguish the phage subsections. Viruses were known to be composed of a protein shell and DNA, so they chose to uniquely label each with a different elemental isotope. This allowed each to be observed and analyzed separately. Since phosphorus is contained in DNA but not amino acids, radioactive phosphorus-32 was used to label the DNA contained in the T2 phage. Radioactive sulfur-35 was used to label the protein sections of the T2 phage, because sulfur is contained in protein but not DNA.
Hershey and Chase inserted the radioactive elements in the bacteriophages by adding the isotopes to separate media within which bacteria were allowed to grow for 4 hours before bacteriophage introduction. When the bacteriophages infected the bacteria, the progeny contained the radioactive isotopes in their structures. This procedure was performed once for the sulfur-labeled phages and once for phosphorus-labeled phages.
The labeled progeny were then allowed to infect unlabeled bacteria. The phage coats remained on the outside of the bacteria, while genetic material entered. Disruption of phage from the bacteria by agitation in a blender followed by centrifugation allowed for the separation of the phage coats from the bacteria. These bacteria were lysed to release phage progeny. The progeny of the phages that were labeled with radioactive phosphorus remained labeled, whereas the progeny of the phages labeled with radioactive sulfur were unlabeled. Thus, the Hershey–Chase experiment helped to confirm that DNA, not protein, is the genetic material.
Confirmation and clarity came a year later in 1953, when James D. Watson and Francis Crick correctly hypothesized, in their journal article "Molecular Structure of Nucleic Acids: A Structure for Deoxyribose Nucleic Acid", the double helix structure of DNA, and suggested the copying mechanism by which DNA functions as hereditary material. Furthermore, Watson and Crick suggested that DNA, the genetic material, is responsible for the synthesis of the thousands of proteins found in cells. They had made this proposal based on the structural similarity that exists between the two macromolecules: both protein and DNA are linear sequences of monomers (amino acids and nucleotides, respectively).
Other experiments
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Once the Hershey–Chase experiment was published, the scientific community generally acknowledged that DNA was the genetic code material. This discovery led to a more detailed investigation of DNA to determine its composition as well as its 3D structure. Using X-ray crystallography, the structure of DNA was discovered by James Watson and Francis Crick with the help of previously documented experimental evidence by Maurice Wilkins and Rosalind Franklin.
Knowledge of the structure of DNA led scientists to examine the nature of genetic coding and, in turn, understand the process of protein synthesis. George Gamow proposed that the genetic code was composed of sequences of three DNA base pairs known as triplets or codons which represent one of the twenty amino acids. Genetic coding helped researchers to understand the mechanism of gene expression, the process by which information from a gene is used in protein synthesis. Since then, much research has been conducted to modulate steps in the gene expression process. These steps include transcription, RNA splicing, translation, and post-translational modification which are used to control the chemical and structural nature of proteins. Moreover, genetic engineering gives engineers the ability to directly manipulate the genetic materials of organisms using recombinant DNA techniques. The first recombinant DNA molecule was created by Paul Berg in 1972 when he combined DNA from the monkey virus SV40 with that of the lambda phage.
Experiments on hereditary material during the time of the Hershey–Chase experiment often used bacteriophages as a model organism. Bacteriophages lend themselves to experiments on hereditary material because they incorporate their genetic material into their host cell's genetic material (making them useful tools), they multiply quickly, and they are easily collected by researchers.
References
External links
- Hershey–Chase experiment animation
- Clear depiction and simple summary
