Recombinant DNA Technology Introduction to Cloning Chromewell Mojica National Institute of Molecular Biology and Biotechnology University of the Philippines, Diliman Quezon City
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Recombinant DNA TechnologyIntroduction to Cloning
Chromewell MojicaNational Institute of Molecular Biology and Biotechnology
University of the Philippines, Diliman Quezon City
At the end of the session, the students should:
1. Be able to understand recombinant DNA technology
2. Be able to define Cloning and the 3 types of cloning
3. Be able to know the different products of recombinant DNA technology
Session Objectives
“Genetic manipulation”
“Genetic modification”
“Recombinant DNA technology”
The technique of altering the genetic makeup of cells or organisms by deliberately inserting, removing, or
altering individual genes
What is Genetic Engineering?
Any organism whose genetic material has been altered.
• Contains a foreign piece of DNA• Deleted some portions of the Genome• Mutated genome
Genetically Modified Organisms (GMOs)
1. Improves crops and livestock2. Massive production of industrially important
enzymes/proteins3. Medical purposes: therapeutic cloning, cheaper
medicine4. Bioremediation5. And MANY more! ☺
The need for Genetic Engineering?
A DNA sequence produced artificially by joining pieces of DNA from different organisms
Created by “cutting and pasting” different DNA fragments
Recombinant DNA (rDNA)
DNA fragment containing the gene ofinterest (transgene)
Plasmid DNA withSelective marker
Restriction enzyme
Ligase
Recombinant DNA (rDNA): The Ingredients
RE: Enzymes that cut DNA in specific sequence
Restriction Enzymes
RE: Enzymes that cut DNA in specific sequence
Recombinant DNA (rDNA): The Ingredients
Production of any organism with the same genetic replica of another cell, tissue or organism
Clone: The organism bearing the copied material
What is Cloning? A clone?
Three Types of Cloning
Similar to recombinant DNA technology
Only certain parts of DNA is copied andtransferred from one organism to another
Uses self replicating PLASMID as VECTOR
DNA/Gene Cloning
Transformation (transfection):Introduction of foreign DNA into cells
Modes of transformation/transfectionElectroporation: Electrical pulseHeat shock: Brief exposure to 37°C-45°CGene gun (biolistics): DNA-coated beads (bullets)Liposome-mediatedViral vectorsInjection of DNA into nucleus
Transformation
Selection and Screening of Successful Transformants
Selection and Screening of Successful Transformants
GM Bacteria for Human Insulin Production
Producing GM Plant
Crown Gall Diseasein Plant
Producing GM Plant
GM Bacteria for Human Insulin Production
Non-Bt corn Bt corn
Three Types of Cloning
Production of a genetic duplicate of an existing organism.
Accomplished using a process called somatic cell nuclear transfer (SCNT)
involves putting the nucleus of a body (somatic) cell into an egg cell from which the nucleus has been removed. (enucleated oocyte)
Reproductive Cloning
Somatic Cell Nuclear Transfer
Other cloned animals via SCNT
Somatic Cell Nuclear Transfer
Three Types of Cloning
Like reproductive cloning but the embryo is not allowed to fully develop
Main purpose is to generate embryonic stem cells (ESC) for therapeutic purposes.
Therapeutic Cloning
STEM CELLS
Mass of undifferentiatedcells that may give rise to different types of cells
Reproductive Cloning
Uses of Stem Cells
Can be used to replace dead cells or damage cells
Can be used to generate specific tissues and organs if provided with specific growth factors
END.
QUESTIONS?
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