HERSHEY-CHASE EXPERIMENT

History Before the Hershey-Chase experiment, proteins were believed to carry the genetic information. At that time, people knew that viruses consisted of DNA (or RNA) inside a protein coat/shell called a capsid. It was also known that viruses replicate by taking over the host cells metabolic functions to make more virus cells. A virus that infects a bacterial cell is called a bacteriophage. A bacteriophage was used in the Hershey-Chase experiment. In order to replicate, the virus must inject the viral genetic code into the host cell. People realized that the viral genetic code material had to be either its DNA or its protein capsid. It was believed to be protein. The results of the Avery-MacLeod-McCarty experiment suggested that DNA was the genetic material, but there was hesitation. The Experiment Hershey and Chase wanted to know whether it was the DNA or the protein coat that was the genetic material. To try to figure this out, Hershey and Chase did an experiment using a bacterium named Escherichia coli, or E. coli and a virus called T2 that is a bacteriophage that infects E. coli. Viruses can only survive on a host cell so T2, like other viruses, must live inside E. coli in order to make more viral cells like itself. When the new T2 viruses are ready to leave the host E. coli cell, they burst the E. coli cell open giving results that the viral DNA, not the protein, is its genetic code material. Hershey and Chase used radioactive chemicals to label the protein capsid and the DNA in T2 virus so they could tell which of those molecules entered the E. coli cells. In the first part of the experiment, the phages were produced in a medium containing 35S radioactively-labeled amino acids. This resulted in proteins being labeled and DNA having no radioactive label. The phage then infected the bacterial cell and injected the DNA. The protein coat remained outside the cell. In the second part the phages were produced in a medium containing 32P labeled deoxyribonucleotides. The DNA was labeled and there was no radioactive label on the protein. Then again the DNA was injected in and the protein coat remained outside. Both parts were then put into a blender to vigorously shake off viral parts stuck on the outer surface of the cell, in this case, protein coat. Then, each mixture was spun in a centrifuge to separate the heavy bacteria (with any viral parts that had gone into them) from the liquid solution they were in (including any viral parts that had not entered the bacteria). The centrifuge causes the heavier bacteria to be pulled to the bottom of the tube where they form a pellet, while the light-weight viral left-overs stay suspended in the liquid portion called the supernatant. The pellet and supernatant from each tube were separated and tested for the presence of radioactivity. Radioactive sulfur was found in the supernatant, indicating that the viral protein did not go into the bacteria. Radioactive phosphorus was found in the bacterial pellet, indicating that viral DNA did go into the bacteria. Conclusion Hershey and Chase concluded that DNA must be the genetic code material, not protein as many poeple believed.