BT 101: Introduction to Biology (3-0-0-6) Semester: 1

(Lectures on Molecular Biology: Dr. Kusum K. Singh)

Course Outline:
DNA
DNA REPLICATION
TRANSCRIPTION
TRANSLATION
CLONING AND
ITS APPLICATION

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 BT 101: Introduction to Biology (3-0-0-6) Semester: 1

Texts:
1. David L. Nelson and Michael M. Cox. Lehninger, Principles of Biochemistry, 6th Ed., Macmillan, Freeman and Company.
2. B. Alberts, A. Johnson, J. Lewis, M. Raff, K. Roberts and P. Walter, Molecular Biology of Cell, 4th Ed., Garland Publishing,
2002.
2. H. Lodish, A. Berk, S. L. Zipursky, M. P. Scott and J. Darnell, Molecular Cell Biology, 4th Ed., W. H. Freeman & Co.,
2003.

References:
1. B. Lewin, Genes VIII, International Edition, Pearson Education, 2004.
2. B. R. Glick and J. J. Pasternak, Molecular Biotechnology: Principles and Applications of Recombinant DNA, 3rd Ed., ASM
Press, 2003.
3. R. M. Twyman, S. B. Primrose and R. W. Old, Principles of Gene Manipulation, Blackwell Science, 2001.

Course Evaluation Policy:

End Semester (09th May 2019): 25 %

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 BT 101: Introduction to Biology (3-0-0-6) Semester: 1

Discussion Timings with Instructor:

1. By prior appointment over email (preferred).

2. Or else, Tuesday between 3 to 4 PM in office (provided no official meetings are scheduled).

3. I will NOT share the slides, please TAKE NOTES!!

Instructor:
Dr. Kusum Kumari Singh
Assistant Professor
Office: O-303, Phone (O): 3206
Email: kusumsingh@iitg.ernet.in
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2‟-Deoxyribose Nucleic Acid

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What is a CRISPR Baby??
Scientist named (He Jiankui)
November 2018
Twin girls- resistant to HIV
Disable CCR5 gene (mutations)
Investigations: Why HIV??
CCR5: impact on cognitive function and intelligence

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Intention:

1. Potential of mol. biology and gene editing

2. Bio-ethics

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When and How the DNA was discovered??

1868 Swiss Chemist Johann Friedrich Miescher

• Discovered DNA
• He isolated something new from the nuclei of eukaryotic cells
• Which he called „nuclein‟ inside the nuclei of human white blood cells
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 2‟-Deoxyribose Nucleic Acid

1881 German biochemist Albrecht Kossel

• Identified nuclein as a nucleic acid.
• He isolated five nucleotide bases: adenine (A), cytosine (C), guanine (G)
• Thymine (T) and Uracil (U) that are building blocks of DNA and RNA.
• Nobel Price: in year 1910
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 2‟-Deoxyribose Nucleic Acid

1919 Russian biochemist Phoebus Levene

• Discovered three major components of nucleotide (phosphate, sugar and base)

• First to discover carbohydrate component of DNA (deoxyribose) and RNA (ribose)
• First to propose „polynucleotide‟ model (yeast nucleic acid)
• Tetranucleotide model G-C-T-A-G-C-T-A 9
 “Transforming Principle”

Focus was epidemiology and

pathology of bacterial pneumonia

1928 British bacteriologist Frederick Griffith

• Studied effects of virulent bacteria vs. nonvirulent bacteria injected into mice.

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 “Transforming Principle”

Focus was epidemiology and

pathology of bacterial pneumonia

1928 British bacteriologist Frederick Griffith

• Studied effects of virulent bacteria vs. nonvirulent bacteria injected into mice.

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Griffith's experiment discovering the "transforming principle"
in pneumococcus bacteria.
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SEARCHING FOR GENETIC MATERIAL

• Oswald Avery, Colin MacLeod, & Maclyn McCarty (1944):

• Reported that “transforming agent” in Griffith's
experiment was DNA.
• Also used the Pneumococcus bacteria and test tubes
(NOT mice)

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DNA is the Molecule of Heredity.
When various isolated chemical
components of the S-strain
pneumococcus cells was mixed the
R-strain pneumococcus cells,
It was shown that the DNA from
the S-strain cells, that caused
transformation.

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 Experiment of Hershey and Chase (1952)

Alfred Hershey and Martha Chase demonstrated the genetic

material is DNA by using viruses that infect bacteria. These
viruses only stay on the outside of the cell when infecting the
cells. Also viruses are composed of protein and DNA. It is
known that the virus injects its genetic material into the
bacterium which had to be either DNA or proteins. 15
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CONCLUSION:
• As Coat of bacteriophage labelled with S35 so the results clearly
indicate that only DNA enter into the host cell & protein coat left
outside.
• The DNA entering the host cell carries all the genetic information
for synthesis of new phage particle.
• This proves that DNA is the genetic material in bacteriophage.

• Nobel Price: in year 1969

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Rosalind Franklin & Maurice Wilkins
Early 1950’s

Rosalind Franklin Maurice Wilkins

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X-ray diffraction showed
•DNA fibers had a twisted pattern
•Large groups of molecules in the fibers
were spaced out in equal intervals

Photo 51
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Linus Pauling James Watson Francis Crick

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James Watson & Francis Crick - 1953

•Watson Crick Model

•“Double Helix”
•Determined the shape and structure
•Adenine bonds with Thymine
•Cytosine bonds with Guanine

• Nobel Prize: in year 1962

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Watson-Crick Model of B-DNA

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 Nucleotides and Nucleosides
• Nucleotide =
– Nitrogeneous base
– Pentose
– Phosphate

• Nucleoside =
– Nitrogeneous base
– Pentose

• Nucleobase =
– Nitrogeneous base

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Nucleobases

• Derivatives of pyrimidine or purine

• Nitrogen-containing heteroaromatic molecules
• Planar or almost planar structures
• Absorb UV light around 250–270 nm

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Pyrimidine Bases

• Cytosine is found in both DNA and RNA

• Thymine is found only in DNA
• Uracil is found only in RNA
• All are good H-bond donors and acceptors
•Neutral molecules at pH 7

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Purine Bases

• Adenine and guanine are found in both RNA

and DNA
• Also good H-bond donors and acceptors
• Neutral molecules at pH 7

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Pentose in Nucleotides

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Pentose in Nucleotides

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 Pentose in Nucleotides
• -D-ribofuranose in RNA
• -2’-deoxy-D-ribofuranose in DNA

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 Pentose in Nucleotides
• -D-ribofuranose in RNA
• -2’-deoxy-D-ribofuranose in DNA

• Different puckered conformations of the sugar ring

are possible

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Pentose in Nucleotides

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Phosphate Group

• Negatively charged at neutral pH

• Typically attached to 5’ position
• Nucleic acids are built using 5’-triphosphates
• ATP, GTP, TTP, CTP
• Nucleic acids contain one phosphate moiety per
nucleotide
• May be attached to other positions

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 Other Nucleotides:
Monophosphate Group in Different Positions

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Phosphate Group

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 -N-Glycosidic Bond
• In nucleotides the pentose ring is attached to the nucleobase via
N-glycosidic bond

• The bond is formed to the anomeric carbon of the sugar in b

configuration
• The bond is formed:
• to position N1 in pyrimidines
• to position N9 in purines
• Bond cleavage is catalyzed by acid

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Conformation around N-Glycosidic Bond

• Relatively free rotation can occur around the N-glycosidic bond

in free nucleotides
• Angle near 0 corresponds to syn conformation
• Angle near 180 corresponds to anti conformation
• Anti conformation is found in normal B-DNA

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 Polynucleotides
• Covalent bonds formed via phosphodiester linkages
• negatively charged backbone
• DNA backbone is fairly stable
• DNA from mammoths?
• Hydrolysis accelerated by enzymes (DNAse)
• RNA backbone is unstable
• In water, RNA lasts for a few years
• In cells, mRNA is degraded in few hours
• Linear polymers
• No branching or cross-links
• Directionality
• 5’ end is different from 3’ end
• We read the sequence from 5’ to 3’
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 Polynucleotides
• Covalent bonds formed via phosphodiester linkages
• negatively charged backbone
• RNA backbone is unstable
• In water, RNA lasts for a few years
• In cells, mRNA is degraded in few hours
• Linear polymers
• No branching or cross-links
• Directionality
• 5’ end is different from 3’ end
• We read the sequence from 5’ to 3’

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 Polynucleotides
• Covalent bonds formed via phosphodiester linkages
• negatively charged backbone
• DNA backbone is fairly stable
• DNA from mammoths?
• Hydrolysis accelerated by enzymes (DNAse)
• RNA backbone is unstable
• In water, RNA lasts for a few years
• In cells, mRNA is degraded in few hours
• Linear polymers
• No branching or cross-links
• Directionality
• 5’ end is different from 3’ end
• We read the sequence from 5’ to 3’
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 Hydrogen-Bonding Interactions
• Two bases can hydrogen bond to form a base pair
• For monomers, large number of base pairs is possible
• In polynucleotide, only few possibilities exist
• Watson-Crick base pairs predominate in double-
stranded DNA
• A pairs with T
• C pairs with G
• Purine pairs with pyrimidine

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DNA_Movie

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Watson-Crick Model of B-DNA

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Other Forms of DNA

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Questions?

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 Questions?
1. How are a nucleoside and a nucleotide similar and how are they different?
2. Match the type of bond with the role below:
Bond-type Role
(a) phosphodiester ___ links base to pentose in nucleotide
(b) N-glycosidic ___ joins adjacent nucleotides in one strand
(c) phosphate ester ___ joins complementary nucleotides in two strands
(d) Hydrogen ___ difference between a nucleoside and a nucleotide
3. Compounds that contain a nitrogenous base, a sugar, and a phosphate group are called (a)________.
Two purines found in DNA are (b)______________ and __________________. A pyrimidine found in all
DNA but not in RNA is (c)_________________. In DNA, the base pair (d)___ –___ is held together by
three hydrogen bonds; the base pair (e)___ –___ has only two such bonds.
4. What is the approximate length of a DNA molecule (in the B form) containing 10,000 base pairs?

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