Historical Background of Molecular
Biology
Frederick Griffith (1928)
•Transformation experiment on Diplococcus
pneumoniae (Pneumococcus)
Smooth type (S)
- thick polysaccharide capsule
- smooth mucoid colonies
- pathogenic / virulent
Rough type (R)
- no capsule
- rough colonies
- non-pathogenic / avirulent
Griffith
ref: Passarge, 2007
Griffith’s Transformation Experiment
Possible Explanations
•R types caused dead S types to come
back to life.
•Information from the heat-killed S type
was transferred to the live R type.
RELATED EXPERIMENTS
M. H. Dawson (1930) and J. L. Alloway
(1932)
in vitro transformation
mouse was not necessary
extract from heat-killed S cells + live R
cells
→live S
CONCLUSION Griffith,Dawson and Alloway
• transformation of the avirulent R type to the
virulent S type
• A transforming principle or agent is
responsible
Avery, MacLeod and McCarty (1944)
Isolation of Transforming Substance
1. Bacterial culture at 37°C for 16-18 hrs
2. Centrifuge
3. Resuspend in 150 ml chilled 0.85 M NaCl
4. Heat kill cells at 65°C and wash with saline
solution
5. Saline + 0.5% Na deoxycholate to extract
water soluble
cell components.
6. Ethanol precipitation
7. Chloroform extraction to remove proteins
and lipids
8. Enzyme digestion to remove polysaccharide
capsule
9. Ethanol precipitation.
a. General Properties of Transforming
Substance
1. viscous, silky sheen
2. loses activity in distilled water
3. activity retained for months in
physiological salt solution
4. withstands heating at 65°C for 30-60
minutes
5. loses activity with increased acid
Concentration
ARGUMENTS against DNA as the genetic
material
DNA is too simple; proteins can account for
more
diversity.
DNA is relevant only to some bacteria; protein
is the more likely genetic material.
The extract was contaminated / not pure
DNA.
Hershey and Chase Blender Experiment
HYPOTHESIS
__________is essential for production of
new phages while__________ are only for
attachment.
Use of T2 Phage:
• Lytic phage
• Lyses cell after 30-60 min
• Releases new phages
Blender Experiment:
• Phages were labeled
• Phages were allowed to infect the cells
• Shearing done using kitchen blender
• Phages were torn off from surface of cells
• Radioactivity measured
1953 --→ 2003 : They took the same picture
again.

PROPERTIES OF GENETIC MATERIAL

The genetic material must be F.A.I.R.S.
• F = flexible/respond to external signals
• A = altered/generate genetic diversity Vitalism
• I = information • Intact cells possess
• R = replicated and transmitted accurately a “vital force”.
• S = stable (very) • To study life, one
must study an intact
Assignment cell.
• On ½ YP, list down the 5 properties of a
genetic material. Using each property as a
Mechanism
heading, enumerate and discuss very
concisely the property/ies of the DNA that • To study life, a cell
makes it a suitable genetic material.
should be dissected
• Please use blue or blank ink only in all
assignments. like a machine.

• Deadline: Next Meeting

Holism
• The whole is greater than the
sum of its parts.

Reductionism
• All biological phenomena can
be reduced into molecules
and atoms→ the laws of
physics and chemistry

MOLECULAR BIOLOGY
BIOCHEMISTRY

Introductory Molecular Biology

Overview
How should we study biological processes? ✓ physical and chemical
Early Schools of Thought: structure of biomolecules
VITALISM vs MECHANISM ✓nucleic acids, proteins,
HOLISM vs REDUCTIONISM lipids, carbohydrates
“STRUCTURAL ROOT” • How is information obtained from DNA?
• How is gene expression regulated?
MICROBIAL GENETICS • How are cellular substances
transported in and out of the cell?

✓information on the genetic • How do specialized cells work?

material, its transmission and muscle cells nerve cells

expression • What is the molecular basis of

✓simple systems, i.e. biological phenomena?

phages, bacteria -Aging, development, immune reaction,

“INFORMATIONAL ROOT” cancer and other human diseases

MOLECULAR BIOLOGY
“the study of the chemical and physical
structure of biological macromolecules”
-Astbury, 1945

2. Physical measurements
Some methods:
a. Ultracentrifugation

MOLECULAR BIOLOGY b. Electrophoresis

(present day definition) c. Infrared absorption spectroscopy

“the study of genes and their

products and how these products 3. In vitro approach

function and interact in the • crude extracts and reconstituted systems

organization and perpetuation of

living things”

PROBLEMS
• What is the genetic material?
• How are macromolecules
proteins) synthesized?
The Logic of MOLECULAR BIOLOGY
(especially
Arguments are based on...

• How does DNA replicate? - Efficiency

- Models
 - Strong inference

Arguments based on:

1.Efficiency
Living cells have had millions of years to
evolve:
Competition and survival selected for
efficiency.
In biological processes, little energy and
material are wasted.
e.g. gene expression

2. Development and evaluation of models

Model
• tentative explanation of how a system works
• tested for validity
The Cell Surface and the Extracellular
Matrix