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Objectives:
- We want to prove the existence of a genetic inheritance in all the alive
beings
- We search to know if the genetic inheritance is identical at all the species.
- We search to know if the genetic inheritance is transmissible in experiments of
a species to another.
Does there exist a genetic
material in all the alive beings?
We wants to extract from the DNA of different cellular
species such as:
- Plant species : the onion
- Animal species eukaryote* pluricellular : saliva
- Animal species eukaryote* pluricellular : liver of mouse
- Animal species prokaryote** : bacterium
*: cell :containing a core
**: cell: not having a core
Material
DNA
human
DNA of
onion
DNA of
liver
DNA of
bacterium
- Saliva
-Onion
- test tube of
50ml
- Plug
(TE*+SDS+Na
Cl)
- A centrifuge
-Alcohol
(ethanol)
-A lump of liver
(2g)
- a Potter
- Tube of 50 ml
- Plug
(TE*+SDS+NaCl
)
-Alcohol
(ethanol)
-Bacterium
- a centrifuge
- Streamroom
- culture
medium
- Plug ( TE*+
SDS+NaCl)
-Alcohol
(ethanol)
- test tube of
15 ml
- Plug (
TE*+SDS+Na
Cl)
- Alcohol
(ethanol)
*: Plug Tris-EDTA, pH 8
Procedure of extraction of DNA
of liver
• LIVER:
• Chop a liver
• Put in a potter and crush
• Put in a plug (SDS+NaCl+
TE) to make burst the cells.
• Centrifuge
• To eliminate the supernatant
• Add the ethanol
• Recover the ball supposed to
be DNA
•
Procedure of extraction of DNA
of bacterium
BACTERIUM:
•
Setting in culture of the
bacterium
•
Put in a tube eppendorf
•
Centrifuge
•
Eliminate the culture medium
•
Put in the plug (SDS+NaCl+TE)
•
Agitate after add ethanol
•
Put in the refrigerator
•
Recover the ball supposed to be
DNA
Procedure of extraction of DNA
of human saliva
HUMAN SALIVA:
• Collect human saliva
• Pour in a tube of 15ml
• Pour the plug (SDS
+NaCl+TE) in this tube
• Pour ethanol
• Let rest
• Recover the ball supposed to
be DNA
Ball of DNA
Procedure of extraction of DNA
of onion
•
ONION
•
Chop the onion
•
Put in a tube of 50mL
•
Put in the plug (SDS+NaCl+TE)
•
Centrifuge
•
Add the ethanol
•
recover the ball supposed to
be DNA
First objective’s conclusion :
We observe a ball, that supposing to
us to be DNA, in the four tubes of the
various cellular species. If that is
checked, we can highlight a genetic
inheritance in the alive beings tested.
Test of characterization of the
DNA
Thanks to the composition of the DNA
that we know, we carried out, in class, an
experiment aiming at proving the presence of
the various elements composing the DNA. A
coloring specific to the DNA consolidated us
in our idea that the collected ball was well of
the DNA.
Is the genetic inheritance
identical between species?
• One treats the DNA with the protein K
"to degrade" proteins related to the
DNA. A fraction of this DNA was
treated with enzymes of restriction.
Analyze by electrophoresis on GEL of agar of DNAs extracted
1
2
3
4
5
6
7 8 9
1 MARKER OF SIZE
2 DNA OF LIVER
3 DNA OF
BACTERIE
4 ONION DNA
5 DNA OF SALIVA
6 DNA OF LIVER
DIGESTED
7 DNA OF BACTERIE
DIGESTED
8 ONION DNA
DIGESTED
9 DNA OF SALIVA
DIGESTED
Second objective’s conclusion:
• One observed thanks to the technique
of electrophoresis that between the
species tested the genetic inheritance
was not identical because the migrations
of the DNA of each species were not
the same ones.
Can one transmit the genetic
inheritance of one species to
the other?
• MATERIALS:
• Bacteria
• - DNA coding the GFP (Fluorescent Green Protein) taken
with the jellyfish
• - box of Petri containing agar
• - Plasmid (circular DNA)
• - Starters
• - Taq DNA Polymerase
• - DNTP
• - Plug for PCR
• - Thermocyclor
• - Tubes of PCR
Technique of the PCR
Séquence cible
GFP
Denaturising of
the DNA
Cycle 1
Hybridizing
Elongation
Cycle 2
Cycle 3
We have to carry out 29 cycles
TRANSFER OF THE GENE OF GFP
RESULTING FROM MEDUSE TO THE
BACTERIE (Escherichia Coli)
GFP
Plasmid
Bacteria
Transformation
(introduction of the plasmid)
Transformed bacterium (GMO)
Obtaining genetically modified
bacteria: fluorescence GFP
Bacteria (control)
Genetically modified bacteria
General conclusion
We have to carry out three
experiments aiming at proving the
existence of the DNA in all the species.
We have deduce the existence from
the DNA, its authenticity between the
species and the possibility of
transmission of the genetic information
from one species to another in
laboratory.
Au vue des résultats de nos
expériences, nous pouvons déduire
qu’ il existe effectivement un
patrimoine génétique chez tous les
êtres vivants testés.