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DNA: The Genetic Material

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DNA: The Genetic Material

Learning Outcomes

 Describe the experiments that first supported the hypothesis that a cell’s hereditary material is
located in the nucleus.
 Understand the theory and conclusions associated with the Griffith and Avery experiments using
Pneumococcus and mice.
 Explain the evidence that supports the identity of DNA as hereditary material.
 Identify the three subunits of DNA and describe how they are put together to construct an intact
molecule.
 Understand the importance of Chargaff’s rules and the complementary nature of nucleotide
bases.
 Describe Watson and Crick’s three--dimensional model of DNA based upon Franklin’s X-ray
crystallography.
 Know what is meant by semi conservative replication of DNA and how it was determined.
 Explain the process of semi discontinuous replication in both strands of the DNA double helix.
 Understand the one gene-one polypeptide hypothesis and the experimental evidence behind it.

HERSHEY AND CHASE EXPERIMENT

 In the Hershey –Chase experiment, bacterial viruses, called phage were used to demonstrate
that DNA is the genetic material. The phage used in this experiment consisted of a DNA
molecule, surrounded by a protein coat.
 When phage infect bacteria, they attach to the surface of the bacterium and inject the DNA into
the cell. The protein coat remains on the outside of the cell.
 In the first part of the experiment, phage were produced in a medium containing 35S
radioactively-labelled amino acids. This resulted in a phage population with 35S-labelled
proteins, but no radioactive label in the DNA.
 The phage were then allowed to infect the bacteria.
 The phage attached to the bacterial cell and injected their DNA, but the radioactively-labeled
protein coat remained on the outside of the cell.
 The phage produced in these cells contained no radioactivity.
 Vigorous shaking caused the empty protein coats to be removed, but did not interfere with
production of new phage in the cell.
 In the second part of the experiment , phage were produced in a medium containing 32P-
labeled deoxyribonucleotides. This resulted in a phage population with 32P-labeled DNA, but no
radioactive label in the protein.
 When the phage infected the bacteria, the 32P-labeled DNA entered the cell and could be found
in phage subsequently produced in the infected bacteria.
This demonstrated that the DNA, but not the protein, carries the genetic information for a new
generation of phage,

MESELSON AND STAHL EXPERIMENT

 The Meselson and Stahl experiment provides evidence for semi-conservative replication of the
DNA molecule where the two parent strands serve as the template for synthesis of new strands.
 In this experiment, bacterial cells were grown for several generations on a medium containing a
heavy isotope of nitrogen(15N). The DNA in these cells therefore contained “heavy” (15N)
nitrogen.
 The cells were then transferred to a new medium containing the normal lighter isotope,(14N).
At various times after the transfer, samples of the bacteria were collected.
 The DNA was then extracted and dissolved in a solution of cesium chloride.
 The samples were then spun rapidly in a centrifuge. When the cesium chloride is centrifuged at
high speed, a concentration gradient is established in the tube.
 DNA molecules move in the gradient until they reach a place where their density equals that of
the cesium.
 DNA containing 14N moved to a position in the gradient determined by its density. DNA
containing 15N is denser than the containing 14N, so it sank to a lower position in the cesium
gradient.
 After one generation in 14N medium, the bacteria yielded a single band of DNA with a density
between that of 14N-DNA, indicating that only one strand of each duplex contained 15N.
 After two generations in 14N medium, two bands were obtained, one of intermediate density
( in which one of the strands contained 15N) and one of low density (in which neither strand
contained 15N)

Meselson and Stahl concluded that replication of the DNA duplex involves building new molecules by
separating parent strands and then adding new nucleotides to form the complementrary strand on each
of these templates.

Griffith and Avery experiments

In the Griffith and Avery experiments, Griffith studied two strains of the bacterium Streptococcus
pneumonia, Bacteria of the S(smooth) strain can cause pneumonia in mice: they are pathogenic because
they have a capsule that protects them from an animal’s defencse system. Bacteria of the R(rough)
strain lack a capsule and are non pathogenic. To test for the trait of pathogenicity, Griffith injected mice
with the two strains as shown below:

Living S cells Mouse dies


Living R cells Mouse healthy
Heat killed S cells Mouse healthy
Mixture of heat killed S cells and living R cells Mouse dies, in blood sample: living S cells are
found that can reproduce, yielding more S cells.
Conclusion, Griffith concluded that the living R bacteria had been transformed into pathogenic S
bacteria by an unknown, heritable substance fron the dead S cells that allowed the R cells to make
capsules.

Membranes
Learning Outcomes

 Understand the biochemistry of phospholipids and how they are organized into membranes.
 Know the function of each of the four components of a cell membrane.
 Differentiate among diffusion, facilitated transport, facilitated diffusion, osmosis, and active
transport.
 Describe the six classes of membrane proteins and how each interacts with the membrane.
 Describe solution and solute movement into and out of a cell under hyperosmotic, hypoosmotic,
or isosmotic conditions.
 Explain and give examples of endocytosis, phagocytosis, pinocytosis, receptor-mediated
endocytosis, and exocytosis.
 Understand the importance of selective permeability in biological systems.
 Describe the operation of the sodium potassium pump and the proton pump.
 Understand the importance of coupled channels, cotransport, and countertransport.

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