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Syllabus

• CBE3014 – Genetic Engineering ( 유전공학 )• Lecture: Mon 13:30 – 14:45

Fri 12:00 – 13:15• Lecture Room: AS301• Instructor: Hyuncheol Kim

R417, hyuncheol@sogang.ac.kr,

705-8922 • Teaching Assistant: Ji-Hwang Lee

R417, leezh44@hanmail.net

010-9666-6726• Grading Policy: Absolute Evaluation

Mid-Term Exam --- 40%

Final Exam --- 40%

Assignment --- 20%

Drug Delivery & Tissue Engineering lab.Sogang University

Syllabus

• Textbook:

Terry A. Brown, Gene Clonging & DNA Analysis (An Introduction), 6th Edition, Wiley-Blackwell Publishing, 2010

Chapter 1. Why Gene Cloning and DNA Analysis are Im-

portant

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What is the genetic engineering?

Genetic engineering is a scientific de-

velopment that involves the artificial ma-

nipulation of an organism’s genes by us-

ing techniques such as molecular cloning

and transformation in order to alter their

nature and structure. Many of these trans-

formations are achieved by manipulation

of an organism’s DNA, which effectively

is the code inscribed in every cell to de-

termine how it will function.

Definition of Genetic engineering

Scientific alteration of the structure of genetic material in a living organism.It involves the production and use of recombinant DNA and has been employedto create bacteria that synthesize insulin and other human proteins.

The Applications of Genetic Engineering

The Applications of Genetic Engineering

Genetech (source : E. coli)

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Genetic Engineering in Medicine

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Genetic Engineering in Agriculture

Engineering resistant corn.

Following the insertion of a

gene from the bacteria Bacillus

thuringiensis, corn becomes re-

sistant to corn borer infection.

This allows farmers to use less

insecticides.

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Genetic Engineering in Energy

Metabolic Engineering

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Genetic Engineering in Energy

The Applications of Genetic Engineering

Gene Therapy

Gene Delivery Vector

The Applications of Genetic Engineering

GM rice delivers antibodies against deadly rotavirus

- Researchers have added an antibody to fight rotavirus into the rice genome.

Researchers engineered the rice, called MucoRice-ARP1, by adding an anti-

body to fight rotavirus originally found in llamas in the rice genome.

Rotavirus is the leading cause of severe diarrhea in young children and in-

fants, killing more than 520,000 people each year, according to the WHO.

More than 85 per cent of those deaths occur in impoverished countries in

Africa and Asia.

The rice could be used to complement vaccinations to protect children when

they are at their most vulnerable to rotavirus, say the researchers.

ZMAPP: EXPERIMENTAL EBOLA TREATMENT PRODUCED IN GENETI-CALLY MODIFIED PLANTS

What is gene cloning ?

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What is gene cloning ?

1. A fragment of DNA, containing the gene to be cloned, is inserted into a circular

DNA molecule called a vector, to produce a recombinant DNA molecule. (Chap-

ter 2, 3, 4)

2. The vector transports the gene into a host cell, which is usually a bacterium, al-

though other types of living cell can be used. (Chapter 5, 7)

3. Within the host cell the vector multiplies, producing numerous identical copies,

not only of itself but also of the gene that it carries. (Chapter 5)

4. When the host cell divides, copies of the recombinant DNA molecule are passed

to the progeny and further vector replication takes place. (Chapter 5)

5. After a large number of cell divisions, a colony, or clone, of identical host cells is

produced. Each cell in the clone contains one or more copies of the recombinant

DNA molecules; the gene carried by the recombinant molecule is now said to be

cloned. (Chapter 6, 8)

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Vector in Molecular Biology

• Plasmid Vector : an extra-chromosomal DNA molecule separate from the choromosomal DNA which is capable of replicating independently of the chromosomal DNA.

• Cloning vector : a small piece of DNA into which a foreign DNA fragment can be inserted.

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Vector in Molecular Biology

• Viral vectors : a tool commonly used by molecular biologists to deliver genetic material into cells. (e.g. Retrovitruses, Lentiviruses, Adenoviruses, Adeno-associated viruses.)

• Non-viral vectors : - Naked DNA - Oligonucleotides - Lipoplexes and polyplexes - Hybrid Methods - Dendrimer - etc.

PCR (Polymerase Chain Reaction)

Definition: A technique to amplify a single or few copies of a piece of DNA across several orders of magnitude, generating millions or more copies of a particular DNA sequence.

Procedure:1. Denaturation of the template DNA - 94oC

2. Annealing of the oligonucleotide primers – 50 ~ 60oC

3. Synthesis of new DNA - 74oC

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PCR

• Summary Movie about PCR (Youtube)

http://www.youtube.com/watch?v=_YgXcJ4n-kQ#watch-main-area

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Why gene cloning and PCR are so important

• Both techniques can provide a pure sample of an in-dividual gene, separated from all the other genes in the cell.

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Obtaining a pure sample of a gene by cloning

Cloning allows individual fragments of DNA to be purified.

The problem of selection

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PCR can also be used to purify a gene

Gene isolation by PCR

The end of Chapter 1