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Griffith Experiment - Transformation in Bacteria, DNA as Genetic Material

Griffith's Experiment in 1928 demonstrated bacterial transformation, where non-virulent bacteria turned virulent upon exposure to heat-killed virulent strains. Avery, MacLeod, and McCarty experiment later confirmed in 1944 that DNA, not proteins, was the genetic material responsible for this transformation. Griffith Experiment in conclusion recognized DNA's significant role in heredity. In this article, we will study the Frederick Griffith Experiment - steps, strain of bacteria, and Griffith Experiment summary.

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Griffith Experiment & Transforming Principle

Griffith experiment diagram, r strain and s strain bacteria.

Griffith’s Experiment - Transformation in Bacteria

Impact of the Griffith Experiment

Dna as genetic material.

Frederick Griffith conducted an experiment that demonstrated the transfer of genetic information between bacteria. The experiment showed that a heat-killed virulent strain could transform a non-lethal strain of bacteria . Griffith called the material that was transferred the "transforming principle". Griffith's experiment involved mixing living non-virulent bacteria with a heat-inactivated virulent form. The bacteria used in the experiment were Streptococcus pneumoniae, which showed two growth patterns. One culture plate had s mooth, shiny colonies (S), while the other had rough colonies (R) .

Griffith's experiment proved that some organisms can acquire new properties from their environment and from one another. However, it took almost 20 years for Avery, McLeod, and McCarty to confirm that nucleic acids, not proteins , are the molecules of heredity

Also Read : Mendel's Laws of Inheritance

The diagram of griffith experiment is shown below:

The R strain and S strain bacteria are two variants of the bacterium Streptococcus pneumonia, used by Frederick Griffith in his experiment. S strains are pathogenic, meaning they can cause disease. R strains are non-pathogenic, meaning they do not cause disease. Some other differences between R and S strains are:

Appearance: S strains have a smooth capsule , or outer coat, made of polysaccharides. R strains lack a capsule and have a rough appearance.
Colonies: S strains produce rough colonies, while R strains produce smooth colonies.
Virulence: S strains are virulent, while R strains are non-virulent.
Immune responses: The capsule of S strains allows the cell to escape the immune responses of the host mouse.
Mice: Mice injected with S strains die within a few days, while mice injected with R strains do not die.

In Griffith's experiment, when he injected mice with the heat-killed S strain and live R strain , the mice unexpectedly died. This revealed a transformation process where the R strain had taken up genetic material from the heat-killed S strain and become virulent. This observation helped in understanding bacterial transformation and the role of DNA as genetic material.

Also Read: Genetic Code - Molecular Basis of Inheritance

Griffith Experiment of Transformation in Bacteria

In 1928, English bacteriologist Frederick Griffith conducted an experiment that demonstrated how bacteria can change their function and form through transformation. The experiment was the first to suggest that bacteria can transfer genetic information through transformation. The experiment involved two strains of the bacterium Streptococcus pneumoniae: a virulent (disease-causing) strain (S) and a non-virulent (non-disease-causing) strain (R).

Transformation is the process of one thing changing into another. In molecular biology and genetics, transformation is the genetic alteration of a cell. It's one of three processes that lead to horizontal gene transfer , along with conjugation and transduction. The detail description of the Griffith’s Experiment - Transformation in Bacteria is as follows:

Griffith Experiment Steps

In the experiment, Griffith injected two types of Streptococcus pneumoniae into mice.

Griffith then subjected the virulent, smooth strain (S) to heat that killed the bacteria. This heat-killed strain (S) was no longer capable of causing disease.
Griffith injected mice with the heat-killed virulent strain (S). Surprisingly, the mice survived, indicating that the heat-killed bacteria alone were not harmful.
Griffith mixed the heat-killed virulent strain (S) with the live non-virulent, rough strain (R) and injected this mixture into mice.
The mice developed pneumonia and died, even though the strain injected was previously non-virulent.

Observations and Conclusion

Griffith concluded that some factor or biomolecule in the heat-killed virulent bacteria (S) had transformed the live non-virulent bacteria (R) into a virulent form. This phenomenon was termed "transformation," though Griffith could not identify the nature of the transforming substance.

Significance

Griffith's experiment laid the groundwork for understanding genetic transformation and proved that DNA , rather than proteins, carried genetic information. This discovery was fundamental to the development of molecular genetics and is also used in a variety of genetic engineering applications.

Conclusion - Griffith Experiment

Frederick Griffith's 1928 experiment on Streptococcus pneumoniae demonstrated bacterial transformation through a transfer of hereditary traits between strains. In Griffith experiment conclusion, the result showed that the harmless R strain could be transformed into a virulent form when exposed to the heat-killed S strain. Subsequent work by Avery, MacLeod, and McCarty in 1944 identified DNA as the transforming substance, establishing it as the genetic material. The discovery laid the foundation for molecular genetics, confirming the role of DNA in transmitting hereditary information.

Also Read: Inherited Traits Lethal Allele - Examples, & its Types Difference Between Phenotype and Genotype Ratio Importance of Variation

FAQs on Frederick Griffith Experiment

What was griffith’s experiment and why was it important.

Frederick Griffith conducted an experiment that suggested bacteria can transfer genetic information through transformation. The experiment was important because it showed that bacteria can change their function and form through transformation.

What is the Griffith Experiment Conclusion?

Frederick Griffith experiment concluded that bacteria can transfer genetic information through a process called transformation.

What was the Most Significant Conclusion of Griffith’s Experiments with Pneumonia in Mice?

Griffith conducted experiments with mice and Streptococcus pneumonia bacteria. He concluded that heat-killed bacteria can convert live avirulent cells to virulent cells. Griffith called this phenomenon transformation.

What did Frederick Griffith Want to Learn about Bacteria?

Frederick Griffith, a British bacteriologist, wanted to learn how bacteria could acquire new traits and how certain types of bacteria produce pneumonia.

How did the Two Types of Bacteria Used by Griffith Differ?

The two types of bacteria used by Griffith were the R strain, lacking a virulent capsule and non-pathogenic, and the S strain, possessing a smooth capsule and causing pneumonia in mice, making it pathogenic.

What was Oswald Avery's Experiment?

The experiment demonstrated that DNA was the only molecule that transformed from one bacterial strain to another.

What is Griffith's Transforming Principle?

Griffith performed an experiment with bacteria and mice and discovered that bacteria can incorporate foreign genetic material from their environment, which he called the transforming principle.

Why is Chapter Griffith Experiment Class 12 Important?

The Griffith Experiment in Class 12 biology is important as it describes bacterial transformation, highlighting the role of genetic material in heredity and laying the foundation for modern molecular biology and genetics research.

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Microbe Notes

DNA Experiments (Griffith & Avery, McCarty, MacLeod & Hershey, Chase)

DNA, deoxyribonucleic acid, is the carrier of all genetic information. It codes genetic information passed on from one generation to another and determines individual attributes like eye color, facial features, etc. Although DNA was first isolated in 1869 by a Swiss scientist, Friedrich Miescher, from nuclei of pus-rich white blood cells (which he called nuclein ), its role in the inheritance of traits wasn’t realized until 1943. Miescher thought that the nuclein, which was slightly acidic and contained a high percentage of phosphorus, lacked the variability to account for its hereditary significance for diversity among organisms. Most of the scientists of his period were convinced by the idea that proteins could be promising candidates for heredity as they were abundant, diverse, and complex molecules, while DNA was supposed to be a boring, repetitive polymer. This notion was put forward as the scientists were aware that genetic information was contained within organic molecules.

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Griffith’s Transformation Experiment

In 1928, a young scientist Frederick Griffith discovered the transforming principle. In 1918, millions of people were killed by the terrible Spanish influenza epidemic, and pneumococcal infections were a common cause of death among influenza-infected patients. This triggered him to study the bacteria Streptococcus pneumoniae and work on designing a vaccine against it . It became evident that bacterial pneumonia was caused by multiple strains of S. pneumoniae, and patients developed antibodies against the particular strain with which they were infected. Hence, serum samples and bacterial isolates used in experiments helped to identify DNA as the hereditary material.

He used two related strains of S. pneumoniae and mice and conducted a series of experiments using them.

When type II R-strain bacteria were grown on a culture plate, they produced rough colonies. They were non-virulent as they lacked an outer polysaccharide coat. Thus, when RII strain bacteria were injected into a mouse, they did not cause any disease and survived.
When type I S-strain bacteria were grown on a culture plate, they produced smooth, glistening, and white colonies. The smooth appearance was apparent due to a polysaccharide coat around them that provided resistance to the host’s immune system. It was virulent and thus, when injected into a mouse, resulted in pneumonia and death.
In 1929, Griffith experimented by injecting mice with heat-killed SI strain (i.e., SI strain bacteria exposed to high temperature ensuing their death). But, this failed to harm the mice, and they survived.
Surprisingly, when he mixed heat-treated SI cells with live RII cells and injected the mixture into the mice, the mice died because of pneumonia. Additionally, when he collected a blood sample from the dead mouse, he found that sample to contain live S-strain bacteria.

Conclusion of Griffith’s Transformation Experiment

Based on the above results, he inferred that something must have been transferred from the heat-treated S strain into non-virulent R strain bacteria that transformed them into smooth coated and virulent bacteria. Thus, the material was referred to as the transforming principle.

Following this, he continued with his research through the 1930s, although he couldn’t make much progress. In 1941, he was hit by a German bomb, and he died.

Avery, McCarty, and MacLeod Experiment

During World War II, in 1943, Oswald Avery, Maclyn McCarty, and Colin MacLeod working at Rockefeller University in New York, dedicated themselves to continuing the work of Griffith in order to determine the biochemical nature of Griffith’s transforming principle in an in vitro system. They used the phenotype of S. pneumoniae cells expressed on blood agar in order to figure out whether transformation had taken place or not, rather than working with mice. The transforming principle was partially purified from the cell extract (i.e., cell-free extract of heat-killed type III S cells) to determine which macromolecule of S cell transformed type II R-strain into the type III S-strain. They demonstrated DNA to be that particular transforming principle.

Initially, type III S cells were heat-killed, and lipids and carbohydrates were removed from the solution.
Secondly, they treated heat-killed S cells with digestive enzymes such as RNases and proteases to degrade RNA and proteins. Subsequently, they also treated it with DNases to digest DNA, each added separately in different tubes.
Eventually, they introduced living type IIR cells mixed with heat-killed IIIS cells onto the culture medium containing antibodies for IIR cells. Antibodies for IIR cells were used to inactivate some IIR cells such that their number doesn’t exceed the count of IIIS cells. that help to provide the distinct phenotypic differences in culture media that contained transformed S strain bacteria.

Observation of Avery, McCarty, and MacLeod Experiment

The culture treated with DNase did not yield transformed type III S strain bacteria which indicated that DNA was the hereditary material responsible for transformation.

Conclusion of Avery, McCarty, and MacLeod Experiment

DNA was found to be the genetic material that was being transferred between cells, not proteins.

Hershey and Chase Experiment

Although Avery and his fellows found that DNA was the hereditary material, the scientists were reluctant to accept the finding. But, not that long afterward, eight years after in 1952, Alfred Hershey and Martha Chase concluded that DNA is the genetic material. Their experimental tool was bacteriophages-viruses that attack bacteria which specifically involved the infection of Escherichia coli with T2 bacteriophage.

T2 virus depends on the host body for its reproduction process. When they find bacteria as a host cell, they adhere to its surface and inject its genetic material into the bacteria. The injected hereditary material hijacks the host’s machinery such that a large number of viral particles are released from them. T2 phage consists of only proteins (on the outer protein coat) and DNA (core) that could be potential genetic material to instruct E. coli to develop its progeny. They experimented to determine whether protein or DNA from the virus entered into the bacteria.

Bacteriophage was allowed to grow on two of the medium: one containing a radioactive isotope of phosphorus( 32 P) and the other containing a radioactive isotope of sulfur ( 35 S).
Phages grown on radioactive phosphorus( 32 P) contained radioactive P labeled DNA (not radioactive protein) as DNA contains phosphorus but not sulfur.
Similarly, the viruses grown in the medium containing radioactive sulfur ( 35 S) contained radioactive 35 S labeled protein (but not radioactive DNA) because sulfur is found in many proteins but is absent from DNA.
E. coli were introduced to be infected by the radioactive phages.
After the progression of infection, the blender was used to remove the remains of phage and phage parts from the outside of the bacteria, followed by centrifugation in order to separate the bacteria from the phage debris.
Centrifugation results in the settling down of heavier particles like bacteria in the form of pellet while those light particles such as medium, phage, and phage parts, etc., float near the top of the tube, called supernatant.

Observation of Hershey and Chase Experiment

On measuring radioactivity in the pellet and supernatant in both media, 32 P was found in large amount in the pellet while 35 S in the supernatant that is pellet contained radioactively P labeled infected bacterial cells and supernatant was enriched with radioactively S labeled phage and phage parts.

Conclusion of Hershey and Chase Experiment

Hershey and Chase deduced that it was DNA, not protein which got injected into host cells, and thus, DNA is the hereditary material that is passed from virus to bacteria.

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