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1. Gene Cloning

2. What is Cloning
• Cloning is the process of producing
individuals with identical or virtually
identical DNA, either naturally or artificially.
In nature, many organisms produce clones
through asexual reproduction.
• Genome Cloning:
• Using nuclear DNA from one organism to
create a second organism with the same
nuclear DNA.

3. Cloning

4. Cloning
• Molecular Cloning:
• The replication of one part of DNA to
produce a population of cells with identical
DNA molecules.
• Cell Based
• PCR

5. What are cloning vectors?
DNA molecules that are used to "transport" cloned
sequences between biological hosts and the test tube.
1. Autonomous Replication.
2. Dominant Genetic marker for selection.
3. Unique Restriction sites.
4. Minimum amount of nonessential DNA to optimize
cloning.

6. Bacterial Plasmids

7. Activity Plan
• 1- Gene of interest
• 2- Preparation of Competent Cells
• 3- Ligation of DNA into Vector
• 4- Transformation
• 5- Growth on Agar plates
• 6- Screening of Interested Gene

9. 1- Restriction Digestion of
DNA

10. Restriction Sites

11. Restriction Digestion of
Plasmid Vector

12. Ligation

13. Transformation
• Exogenous DNA into cells (E. coli)
• Two methods of Transformation
a- Chemical method using CaCl2
B- Electroporation

14. Transformation: CaCl2

15. Transformation: Electroporation

16. What did the cells take up?

17. Insulin Production by Cloning