ứng dụng công nghệ gen trong y học và chẩn đoán

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+ A,B, C sections of
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Signal sequence is
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to RER
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RER (forming
proinsulin)
Removal of 33 amino
acids at Golgi
and S-S joining of A
and B chains to form
insulin

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Insulin is made up of 2 chains = 51 amino acids total
A chain = 21 amino acids
B chain = 30 amino acids
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Human Insulin Production by Bacteria
6) join the plasmid and human fragment
and cut with a restriction enzyme
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Human Insulin Production by Bacteria
Mix the recombinant plasmid with
bacteria.
Screening bacterial cells to learn which contain the human insulin gene is the
hard part.
 
Route to the Production by Bacteria of Human Insulin
A fermentor used to grow
recombinant bacteria.
This is the step when gene cloning takes place.
The single recombinant plasmid replicates within a cell.
Then the single cell with many recombinant plasmids produces trillions of
like cells with recombinant plasmid – and the human insulin gene.
One cell with the
recombinant plasmid
 
Route to the Production by Bacteria of Human Insulin

The final steps are to collect the bacteria, break open the cells, and purify the
insulin protein expressed from the recombinant human insulin gene.
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Route to the
Production by
Bacteria of
Human Insulin
Overview of gene
cloning.
Cloning animati
on
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If adult suffers with hypopituitarism,
HGH therapy is also available
as normally payable under an insurance policy.
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6./V)4'9/-
HGH could be purified from cadaver (tử thi) pituitary glands
k4HVHO,.0’J,H.y/.k‚;vJ,H.06,.6H'9,-'
Creutzfeldt-Jacob Disease (CJD) is one of the risk of using brain tissue
24 cases reported by 1993 in France from cadaver HGH
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Native human GH contains signal peptide
needed to direct nascent polypeptide to RER in
eukaryotes

Signal peptide is bad for E.coli
(it can not synthetize this protein)
Signal sequence removed be EcoRI cleavage
Coding sequence for first 24 aa also lost in the
process of removal
24 amino acids added back by synthetic DNA linker
Expressed in E. coli successfully
hGH therapy is very expensive, costing anywhere from $800 to $2,500 a month
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