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12TH. CH.4-Molecular Basis
of Inheritance
4.1 The Discovery of DNA:
In 1869, Friedrich Miescher From the pus
cell nuclei, he isolated a unique chemical
substance to which he called nuclein.
Chemically, nuclein has high phosphorus
content.
Moreover it showed acidic properties.
Hence it was named as nucleic acid.
By the early 1900s, we knew that
Miescher's nuclein was a mix (mixture) of
proteins and nucleic acids. There are two
kinds
of nucleic acids. DNA (deoxyribonucleic
4.2 The Genetic Material is a DNA:
By the early 1900s, geneticist knew that
genes control the inheritance of traits, that
genes are located on chromosome and that
chemically chromosomes are mainly
composed
of DNA and proteins.
protein DNA
large small
complex molecules simple molecule.
belived that it store
information needed to
govern cell metabolism.
Hence it was assumed
that proteins caused the
variations observed
within species.
simple molecule whose
composition
varied little among
species.
so it is not surprising that most
researchers initially favored proteins as
the genetic material.
Initially, most geneticists thought that protein
are large, complex molecules and store
information needed to govern cell metabolism.
Hence it was assumed that proteins caused the
variations observed within species.
On the other hand DNA thought to be
small, simple molecule whose composition
varied little among species.
Over the time, these ideas about DNA were shown to
be
wrong. In fact DNA molecules are large and vary
tremendously within and among species.
Variations in the DNA molecules are different than
the variation in shape, electrical charge and function
shown by proteins
Over a period of roughly 25 years (1928-
1952), geneticists became convinced that DNA
and not protein, was the genetic material. Let
us study three important contributions that
helped cause this shift of opinion.
Griffith’s experiments :
In 1928, a British medical officer Frederick
Griffith performed an experiment on bacterium
Streptococcus pneumoniae that causes
pneumonia in humans and other mammals.
Griffith used two strains or two genetic varieties
of Streptococcus to find a cure for pneumonia,
which was a common cause of death at that
time.
The two strains used were :
i. Virulent, smooth, pathogenic and
encapsulated S type.
ii. Non-virulent, rough, non-pathogenic and
non-capsulated R type.
Griffith conducted four experiments on
these bacteria.
1-First, when he injected bacteria of strain R to mice,
the mice survived because it did not develop
pneumonia.
2-Second, when he injected bacteria of strain S to
mice, the mice developed pneumonia and died.
3-In the third experiment, he injected heat-killed strai
S
bacteria to mice, once again the mice survived.
4-In fourth experiment, he mixed heat-killed S bacter
with live bacteria of strain R and injected to mice. Th
mice died and Griffithrecovered large numbers of liv
strain S
bacteria from the blood of the dead mice.
In these four experiments, something had
caused harmless strain R bacterium to change
into deadly S strain bacterium. Griffith showed
that the change was genetic.
He suggested that genetic material from heat-killed
strain
S bacterium had somehow changed the living
strain R bacterium into strain S bacterium.
Griffith concluded that the R-strain
bacterium must have taken up, to what he
called a "transforming principle" from the heat-
killed S bacterium, which allowed R strain to
get transformed into smooth-coated bacterium
and become virulent.
Avery, McCarty and MacLeod’s experiment:
In 1944, after some 10 years of research
and experimentation, U. S. microbiologists
Oswald T. Avery, Colin M. MacLeod and
Maclyn McCarty (all at Rockefeller University
in New York) first evidenced to prove the DNA
is a genetic material (transforming principle),
through the experiments
. They purified DNA, RNA, Proteins (enzymes) and
other materials from cell free extract of S cells/
strain and mixed
with heat killed S strain and R cells seperately
to confirm which one could transform living
R cells into S cells.
Only DNA was able to transform harmless strain R
into deadly strain S.
They also discovered that protein –digesting
enzymes (proteases), RNA-digesting enzyme
(RNAases) did not affect transformation, so the
transforming substance was neither a protein
nor RNA.
DNA digested with DNAse did inhibit the
transformation, suggesting that DNA caused the
. These experiments proved that the transforming
principle is DNAbut all biologists were not
convinced.
Finally, Alfred Hershey and Martha Chase
(1952) proved that DNA is the genetic material
and not proteins, by using bacteriophages.
Hershey - Chase Experiment:
Hershey and Chase worked with viruses
that infect bacteria i.e. bacteriophages, which
are composed of DNA and protein.
They used radioactive phosphorous 32P in the
medium for
some viruses and radioactive sulphur 35S for
some others.
They grew some viruses on a medium that
contained radioactive phosphorus and some
others on medium that contained radioactive
sulphur.
Viruses grown in the presence of
radioactive phosphorus contained radioactive
DNA (labelled DNA), but not radioactive
proteins because DNA contains phosphorus
(labelled DNA) but proteins do not.
Similarly,viruses grown on radioactive sulphur
contained radioactive protein but not
radioactive DNA
because DNA does not contain sulphur.
Radioactive phages were allowed to
infect E.coli bacteria grown on the medium
containing normal ‘P’ and ‘S’. Then, as the
infection proceeded, the viral coats were
removed with the help of centrifuge.
Bacteria which were infected by viruses with
radioactive DNA, were radioactive, indicating
that DNA was the material that passed from
the viruses to the bacteria.
Bacteria which were infected by viruses having
radioactive sulphur (protein) were not
radioactive.
This indicates that proteins from the viruses, did
not enter the bacteria.
DNA is, therefore, the genetic
material that is passed from virus to bacteria
In other words, sometime after infection,
radioactivity for ‘P’ and ‘S’ was tested.
Only radioactive ‘P’ was found inside the bacterial
cell, indicating that DNA is the genetic material.
MOLECULAR BASIS OF INHERITANCE -DNA AS GENETIC MATERIAL
MOLECULAR BASIS OF INHERITANCE -DNA AS GENETIC MATERIAL
MOLECULAR BASIS OF INHERITANCE -DNA AS GENETIC MATERIAL
MOLECULAR BASIS OF INHERITANCE -DNA AS GENETIC MATERIAL
MOLECULAR BASIS OF INHERITANCE -DNA AS GENETIC MATERIAL

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MOLECULAR BASIS OF INHERITANCE -DNA AS GENETIC MATERIAL

  • 2. 4.1 The Discovery of DNA: In 1869, Friedrich Miescher From the pus cell nuclei, he isolated a unique chemical substance to which he called nuclein. Chemically, nuclein has high phosphorus content. Moreover it showed acidic properties. Hence it was named as nucleic acid. By the early 1900s, we knew that Miescher's nuclein was a mix (mixture) of proteins and nucleic acids. There are two kinds of nucleic acids. DNA (deoxyribonucleic
  • 3.
  • 4.
  • 5. 4.2 The Genetic Material is a DNA: By the early 1900s, geneticist knew that genes control the inheritance of traits, that genes are located on chromosome and that chemically chromosomes are mainly composed of DNA and proteins.
  • 6. protein DNA large small complex molecules simple molecule. belived that it store information needed to govern cell metabolism. Hence it was assumed that proteins caused the variations observed within species. simple molecule whose composition varied little among species. so it is not surprising that most researchers initially favored proteins as the genetic material.
  • 7. Initially, most geneticists thought that protein are large, complex molecules and store information needed to govern cell metabolism. Hence it was assumed that proteins caused the variations observed within species. On the other hand DNA thought to be small, simple molecule whose composition varied little among species.
  • 8. Over the time, these ideas about DNA were shown to be wrong. In fact DNA molecules are large and vary tremendously within and among species. Variations in the DNA molecules are different than the variation in shape, electrical charge and function shown by proteins
  • 9. Over a period of roughly 25 years (1928- 1952), geneticists became convinced that DNA and not protein, was the genetic material. Let us study three important contributions that helped cause this shift of opinion. Griffith’s experiments : In 1928, a British medical officer Frederick Griffith performed an experiment on bacterium Streptococcus pneumoniae that causes pneumonia in humans and other mammals. Griffith used two strains or two genetic varieties of Streptococcus to find a cure for pneumonia, which was a common cause of death at that time.
  • 10. The two strains used were : i. Virulent, smooth, pathogenic and encapsulated S type. ii. Non-virulent, rough, non-pathogenic and non-capsulated R type.
  • 11. Griffith conducted four experiments on these bacteria. 1-First, when he injected bacteria of strain R to mice, the mice survived because it did not develop pneumonia. 2-Second, when he injected bacteria of strain S to mice, the mice developed pneumonia and died. 3-In the third experiment, he injected heat-killed strai S bacteria to mice, once again the mice survived. 4-In fourth experiment, he mixed heat-killed S bacter with live bacteria of strain R and injected to mice. Th mice died and Griffithrecovered large numbers of liv strain S bacteria from the blood of the dead mice.
  • 12.
  • 13.
  • 14.
  • 15. In these four experiments, something had caused harmless strain R bacterium to change into deadly S strain bacterium. Griffith showed that the change was genetic. He suggested that genetic material from heat-killed strain S bacterium had somehow changed the living strain R bacterium into strain S bacterium. Griffith concluded that the R-strain bacterium must have taken up, to what he called a "transforming principle" from the heat- killed S bacterium, which allowed R strain to get transformed into smooth-coated bacterium and become virulent.
  • 16. Avery, McCarty and MacLeod’s experiment: In 1944, after some 10 years of research and experimentation, U. S. microbiologists Oswald T. Avery, Colin M. MacLeod and Maclyn McCarty (all at Rockefeller University in New York) first evidenced to prove the DNA is a genetic material (transforming principle), through the experiments
  • 17.
  • 18.
  • 19. . They purified DNA, RNA, Proteins (enzymes) and other materials from cell free extract of S cells/ strain and mixed with heat killed S strain and R cells seperately to confirm which one could transform living R cells into S cells. Only DNA was able to transform harmless strain R into deadly strain S. They also discovered that protein –digesting enzymes (proteases), RNA-digesting enzyme (RNAases) did not affect transformation, so the transforming substance was neither a protein nor RNA. DNA digested with DNAse did inhibit the transformation, suggesting that DNA caused the
  • 20. . These experiments proved that the transforming principle is DNAbut all biologists were not convinced. Finally, Alfred Hershey and Martha Chase (1952) proved that DNA is the genetic material and not proteins, by using bacteriophages.
  • 21.
  • 22. Hershey - Chase Experiment: Hershey and Chase worked with viruses that infect bacteria i.e. bacteriophages, which are composed of DNA and protein. They used radioactive phosphorous 32P in the medium for some viruses and radioactive sulphur 35S for some others. They grew some viruses on a medium that contained radioactive phosphorus and some others on medium that contained radioactive sulphur.
  • 23. Viruses grown in the presence of radioactive phosphorus contained radioactive DNA (labelled DNA), but not radioactive proteins because DNA contains phosphorus (labelled DNA) but proteins do not. Similarly,viruses grown on radioactive sulphur contained radioactive protein but not radioactive DNA because DNA does not contain sulphur.
  • 24. Radioactive phages were allowed to infect E.coli bacteria grown on the medium containing normal ‘P’ and ‘S’. Then, as the infection proceeded, the viral coats were removed with the help of centrifuge. Bacteria which were infected by viruses with radioactive DNA, were radioactive, indicating that DNA was the material that passed from the viruses to the bacteria. Bacteria which were infected by viruses having radioactive sulphur (protein) were not radioactive.
  • 25. This indicates that proteins from the viruses, did not enter the bacteria. DNA is, therefore, the genetic material that is passed from virus to bacteria In other words, sometime after infection, radioactivity for ‘P’ and ‘S’ was tested. Only radioactive ‘P’ was found inside the bacterial cell, indicating that DNA is the genetic material.