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Side by side experiments are performed with separate bacteriophage (virus) cultures in which either the protein capsule is labeled with radioactive sulfur or the DNA core is labeled with radioactive phosphorus. 1. The radioactively labeled phages are allowed to infect bacteria. 2. Agitation in a blender dislodges phage particles from bacterial cells. 3. Centrifugation concentrates cells, separating them from the phage particles left in the supernatant. Results: 1. Radioactive sulfur is found predominantly in the supernatant. 2. Radioactive phosphorus is found predominantly in the cell fraction, from which a new generation of infective phage can be isolated.
which had first been demonstrated in the 1944 Avery±MacLeod±McCarty experiment. coli. After separation.php The Hershey±Chase experiments were a series of experiments conducted in 1952 by Alfred Hershey and Martha Chase.´ http://en. and through several elegant experiments were able to observe the transfer of P32 labeled phage DNA into the cytoplasm of the bacterium.accessexcellence. the radioactive S35 tracer was observed in the protein shells. Following infection of E. but not in the infected bacteria.org/RC/VL/GG/hershey. most assumed at the time that proteins carried the information for inheritance. The phage infects a bacterium by attaching to its outer membrane and injecting its genetic material and leaving its empty shell attached to the bacterium. Hershey and Chase conducted their experiments on the T2 phage. confirming that DNA was the genetic material. they labeled the phages with radioactive Sulfur-35 (Sulfur is present in the amino acids cysteine and methionine. a virus whose structure had recently been shown by electron microscopy. but not in DNA). Structural overview of T2 phage In their first set of experiments. While DNA had been known to biologists since 1869. supporting the hypothesis that the genetic material which infects the bacteria was DNA and not protein.The phage consists of a protein shell containing its genetic material. In their second set of experiments.Conclusion: The active component of the bacteriophage that transmits the infective characteristic is the DNA. Hershey and Chase labeled the DNA of phages with radioactive Phosphorus-32 (the element phosphorus is present in DNA but not present in any of the 20 amino acids from which proteins are made).org/wiki/Hershey%E2%80%93Chase_experiment . Hershey shared the 1969 Nobel Prize in Physiology or Medicine for his ³discoveries concerning the genetic structure of viruses. They allowed the phages to infect E.wikipedia. coli they then sheared the viral protein shells off of infected cells using a high-speed blender and separated the cells and viral coats by using a centrifuge. There is a clear correlation between DNA and genetic information http://www.
The phage used consisted of a DNA molecule. with their DNA going to the inside. or E. coli with a source of radioactive phosphorus. A virus which infects a bacterium is called abacteriophage because the host bacterium cell is killed as the new virus particles leave the bacterial cell. This resulted in phages with the S labeled proteins.swf In 1952. the virus must inject whatever is the viral genetic code into the host cell. Hershey and Chase grew two batches of T2 and E. When the new T2 viruses are ready to leave the host E. is just a crystal of DNA and protein. coli cell open. http://highered. coli in order to make more virus like itself. The protein coat remains on the outside of the cell. like other viruses. The phages infected the bacteria and the phages produced in the cells contained the radioactivity. It was also known that viruses replicate by taking over the host cell s metabolic functions to make more virus. Click on the virus button to study their experiment. First.com/olc/dl/120076/bio21. it has been nicknamed the Hershey-Chase Experiment . called phage. Hershey and Chase sought an answer to the question. killing it (hence the name bacteriophage ). different kinds of viruses that infect other kinds of animals. This was to give the T2 time to inject their genetic material into the bacteria. still other viruses which infect plants. coli with a source of radioactive sulfur. not the protein. Isolated T2. coli cell (and go infect others). coli for short (named after a scientist whose last name was Escher) and a virus called T2 that is a bacteriophage that infects E. We are used to thinking and talking about viruses which invade our bodies and make us sick. the DNA. new batches of E. making that radioactive.In the experiment. these radioactive T2 were placed in separate. Thus. Alfred Hershey and Martha Chase did an experiment which is so significant. The results that Hershey and Chase obtained indicated that the viral DNA. but no radioactive label in the protein. These phage infected bacteria. In the . but were left there only 10 minutes. Since DNA has lots of phosphorus in its phosphate ( PO4) groups.mcgraw-hill. The phage produced in the cells contained no radioactivity but the phage protein coat did. so it must live inside E. Second. people knew that viruses were composed of DNA (or RNA) inside a protein coat/shell called a capsid. Hershey and Chase performed an experiment using a bacterium named Escherichia coli. At that time. Since some amino acids contain sulfur in their side chains. carries the genetic information for a new generation of phage. the phage were produced in a medium containing 35S radioactively-labeled amino acids. surrounded by a protein coat. This resulted in phages with P labeled DNA. In order to do all this. coli. but not the protein. if T2 is grown in E. if T2 is grown in E. but no radioactive label in the DNA. they burst the E. people realized that the viral genetic code material had to be either its DNA or its protein capsid. coli cells. Is it the viral DNA or viral protein coat (capsid) that is the viral genetic code material which gets injected into a host bacterium cell? To try to answer this question. bacterial viruses. Then. but not reproduce. but there are other. coli: one with radioactive sulfur and one with radioactive phosphorus to get batches of T2 labeled with either radioactive S or radioactive P. Hershey and Chase used radioactive chemicals to distinguish between ( label ) the protein capsid and the DNA in T2 virus so they could tell which of those molecules entered the E. phage were produced in a medium containing 32P labeled deoxyribonuleotides. and even some viruses that infect bacteria. Thus. the sulfur will be incorporated into the T2 protein coat making it radioactive. coli. is its genetic code material. were used to demonstrate that DNA is the genetic material. the phosphorus will be incorporated into the viral DNA.
indicating that the viral protein did not go into the bacteria.next step. still in separate batches.uc. indicating that viral DNA did go into the bacteria. there was a lot of competition to be the first to discover its chemical structure. In the subsequent step. coli but perhaps stuck on the outer surface. this would differentiate between the protein and DNA portions of the virus. Hopefully. Hershey and Chase concluded that DNA must be the genetic code material. http://biology. Then.htm . The centrifuge causes the heavier bacteria to be pulled to the bottom of the tube where they form a pellet. Based on these results.clc. Radioactive sufur was found in the supernatant. the pellet and supernatant from each tube were separated and tested for the presence of radioactivity. When their experiment was published and people finally acknowledged that DNA was the genetic material. the mixtures were agitated in a kitchen blender to knock loose any viral parts not inside the E. Radioactive phosphorus was found in the bacterial pellet. not protein as many poeple believed. 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).edu/courses/bio104/dna. while the light-weight viral left-overs stay suspended in the liquid portion called the supernatant.