Historical Background of

Molecular Biology

NO Villa, UPLB
JRA Gomez, MSUIIT
NO Villa, UPLB
JRA Gomez, MSUIIT
Frederick Griffith (1928)
•Transformation experiment on Diplococcus
pneumoniae (Pneumococcus)

Smooth type (S) Rough type (R)

- thick polysaccharide capsule - no capsule
- smooth mucoid colonies - rough colonies
- pathogenic / virulent - non-pathogenic / avirulent

NO Villa, UPLB
JRA Gomez, MSUIIT
Griffith
ref: Passarge, 2007
NO Villa, UPLB
JRA Gomez, MSUIIT
Griffith
 Griffith’s Transformation Experiment
Results
D. pneumoniae Injected
Mice Cells Recovered
Live S Dead S
Live R Alive None
Heat-killed S Alive None
Heat-killed S, Live R Dead S

NO Villa, UPLB
JRA Gomez, MSUIIT
Griffith
 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.

NO Villa, UPLB
JRA Gomez, MSUIIT
Griffith
 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
NO Villa, UPLB
JRA Gomez, MSUIIT
Dawson and Alloway
ref: Passarge, 2007 NO Villa, UPLB
JRA Gomez, MSUIIT
Dawson and Alloway
 CONCLUSION

• transformation of the avirulent R type to

the virulent S type
• A transforming principle or agent is
responsible

NO Villa, UPLB
JRA Gomez, MSUIIT
Griffith,Dawson and Alloway
NO Villa, UPLB
JRA Gomez, MSUIIT
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
NO Villa, UPLB
9. Ethanol precipitation. JRA Gomez, MSUIIT
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
NO Villa, UPLB
JRA Gomez, MSUIIT

Avery, Mcleod and McCarty

e. Enzymatic Analysis
Enzyme Effect
Proteases e.g. trypsin, chymotrypsin Negative
Lipase Negative
RNAse Negative
DNase
Destroys activity
heatedDNase Negative

NO Villa, UPLB
JRA Gomez, MSUIIT
NO Villa, UPLB
JRA Gomez, MSUIIT
 (1869)
“a new acidic, phosphorus-containing,
long molecule”

NO Villa, UPLB
JRA Gomez, MSUIIT
 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.

NO Villa, UPLB
JRA Gomez, MSUIIT
NO Villa, UPLB
JRA Gomez, MSUIIT
Hershey and Chase Blender Experiment

HYPOTHESIS

is essential for production of

new phages while
are only for attachment.

NO Villa, UPLB
JRA Gomez, MSUIIT
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 NO Villa, UPLB
22
JRA Gomez, MSUIIT
NO Villa, UPLB
JRA Gomez, MSUIIT
James Watson and Francis Crick
(1953)
Watson (1928-

Crick (1916-2004)
)

NO Villa, UPLB
JRA Gomez, MSUIIT
 crucial data from other researches…
1. ____ _

DNA is made of nucleotides (phosphate,

sugar, ATCG) -

NO Villa, UPLB
JRA Gomez, MSUIIT
2.

1 purine :
1 pyrimidine
NO Villa, UPLB
JRA Gomez, MSUIIT
3.
- suggested that DNA is triple-stranded

4.
- provided X-ray diffraction photograph of
crystalline DNA.

NO Villa, UPLB
JRA Gomez, MSUIIT
NO Villa, UPLB
JRA Gomez, MSUIIT 28
NO Villa, UPLB
JRA Gomez, MSUIIT
Photo: Cold Spring Harbor Laboratory Archives
32
1953 2003

NO Villa, UPLB
JRA Gomez, MSUIIT
 PROPERTIES OF
GENETIC
MATERIAL

The genetic material must be F.A.I.R.S.

NO Villa, UPLB
JRA Gomez, MSUIIT
The genetic material should be F.A.I.R.S.

• F = flexible/respond to external signals

• A = altered/generate genetic
diversity
• I = information
• R = replicated and transmitted accurately
• S = stable (very)

NO Villa, UPLB
JRA Gomez, MSUIIT
 Assignment
• On ½ YP, list down the 5 properties of a
genetic material. Using each property as a
heading, enumerate and discuss very
concisely the property/ies of the DNA that
makes it a suitable genetic material.
• Please use blue or blank ink only in all
assignments.
• Deadline: Next Meeting
NO Villa, UPLB
JRA Gomez, MSUIIT