wide production of insulin by using E.coli

2. What is meant by Recombinant DNA?
• DNA from DNA from organism B
• organism A
•
• E.g. ;Human e.g.; Bacteria

3. What is meant by cloning ?
• Cutting gene or apiece of DNA from an
organism by using restriction enzymes and
sticking it to DNA of anther organism throw a
vector (usually plasmid or phage )by using
DNA ligase

4. Cloning process
depends on;
• Restriction enzymes cut the gene
• Ligase enzyme stuck the gene to
the vector

5. Firstly ;restriction enzymes
• Restriction enzymes are very specific and
unique for DNA sequence so we use the same
restriction enzyme to cut the DNA sequence
&the plasmid

6. • The bp at which the restriction enzyme
starts&ends called sticky ends because there
is one sequence of DNA That can stick here

8. Production of human insulin by using
genetic engineering
• The diabetes happens because of the
decrease in insulin conc in blood
• The insulin produced from B-cells that found
in pancreas
• At the first :diabetes was treated by using
animals(pigs)insulin but its high cost &side
effects encourage the scientists to think about
using cloned insulin

9. • The insulin that secreted from B-cells isn't in
active form but it must pass by three stages ;
• 1-preproinsulin
• 2-proinsulin
• 3-active insulin

10. • 1-preproinsulin :
• Consists of ,
• 1-N-terminal 16aminoacid
• 2-C-peptide 32amino acid
• 2-A-chain 21aminoacid
• 4-B-chain 30aminoacid

12. • 2-pro insulin ;
• Consists of
• 1-A-chain
• 2-B-chain
• 3-C-peptide
• 3-insulin ;
• 1-A-chain
• 2-B-chain

13. Cloning of active
insulin
Cloning gene A in a
species
Of E.COLI
Cloning gene B in
anther species of
E.COLI

14. Where I will but the gene A or B on the
genome ????
• The E.COLI has the ability to analyze the
lactose sugar by using B-galactosidase
(that isn't found in anther type of bacteria )
• So the genome of the bacteria have lac
operon

15. • #selection of cloned bacteria for A or B genes
by using amp (anti biotic)
• extract A or B from E.COLI
• Mix the 2 poly peptide with each other to form
the active insulin