Professor
• Name: Hyuncheol Kim
• Office address: Engineering Building (Bldg R) Rm 503A
• E-mail: hyuncheol@[Link]
• Phone #: 02-705-8922
• Name of Lab: Functional Nanomedicine Laboratory
기능성 나노의약 연구실
• Homepage: [Link]
• Office Hour: 화요일, 수요일, 목요일 10:00 – 12:00
✓ 면담은 이메일 또는 saint 에서 면담 신청을 하는 것으로 권
장합니다.
Course Overview
• Course Format
- Lecture (70%) & Discussion (30%)
• Evaluation
- Mid-Term Exam : Chapter 1 – 9 (30%)
- Final Exam (30%) : Chapter 10 – End
- Project (30%)
- 출결 (10%)
- I will often introduce the latest biotechnology.
I plan to make exam questions using contents related to the
latest biotechnology.
Main Textbook
Gene Cloning and DNA analysis: An Introduction, T.A
Brown, 6ed, Wiley-Blackwell publishing,
2021
Course Overview
Genetic engineering is a technology that allows scientists to
manipulate the genes of living organisms to express desired traits.
This semester's genetic engineering class will consist of
understanding the principles of gene cloning and producing
recombinant DNA using genetic engineering technology, as well as
introducing various applications using the latest genetic
engineering technology. In addition, during the class, I will have
time to briefly introduce theories on the latest biotechnology
development trends as well as genetic engineering.
Synthetic Biology
Prokaryotic vs. Eukaryotic cells
Prokaryotes are organisms made up of cells that lack a cell nucleus or any membrane-
encased organelles. Eukaryotes are organisms made up of cells that possess a
membrane-bound nucleus that holds genetic material as well as membrane-bound
organelles.
Prokaryotic vs. Eukaryotic cells
Plasmid : Circular, double stranded, supercoiled gene structure
Plasmid is a non-chromosomal circular DNA molecule that exists independently of
chromosomes in bacterial cells and can independently replicate/proliferate.
Coloured TEM of a plasmid of bacterial
DNA - Stock Image - G110/0188 -
Science Photo Library
Bacterium
What is 1 Gene inserted into
plasmid
Cell containing gene
of interest
gene cloning? Bacterial Plasmid
chromosome Gene of
Recombinant interest DNA of
DNA (plasmid)
2 Plasmid put into chromosome
bacterial cell (“foreign” DNA)
Recombinant
bacterium
3 Host cell grown in culture to
form a clone of cells containing
the “cloned” gene of interest
Gene of Protein expressed from
interest gene of interest
Copies of gene Protein harvested
4 Basic research
Basic and various Basic
research applications research
on gene on protein
Gene for pest Gene used to alter Protein dissolves Human growth
resistance inserted bacteria for cleaning blood clots in heart hormone treats
into plants up toxic waste attack therapy stunted growth
The purpose of gene cloning is to clone a
small number of genes of interest to secure a
sufficient number of genes of interest for
basic research and research on proteins
expressed by the genes.
What is PCR (Polymerase Chain Reaction)?
Procedure of 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
Another application of PCR (RT-PCR)
• For amplifying DNA from RNA
• Reverse transcriptase reverse
transcribes RNA into cDNA,
which is then amplified by PCR.
Another application of PCR (RT-PCR and real time - PCR)
MCF-7
10000 MCF-7/ADR
Techniques to compare
Relative mRNA expression
5000
interested gene expression
1 between groups (different cells
or tissues) qualitatively and
0
ABCB1 ABCC1 ABCG2 quantitatively
Importance of PCR (Polymerase Chain Reaction)
Obtaining a pure sample PCR can also be used
of a gene by cloning to purify a gene
Why gene cloning and PCR are so important
Both techniques (Gene Cloning & PCR)
can provide a pure sample of an individual
gene, separated from all the other genes in
the cell.
What will we learn about 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.
(Chapter 2, 3, 4)
2. The vector transports the gene into a host cell, which is usually a bacterium,
although 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. 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)
Organoid
Organoid are in vitro generated three-dimensional (3D), mini cluster of cells that highly
simulate the structure and function of the corresponding organ in vivo.
In 2009, Hans Clevers and colleagues first demonstrated that single Lgr5+ expressing
intestinal adult stem cells (ASCs) could self-organize and differentiate to form intestinal
crypt-villus structures that contained all intestinal cell types. In the following decades,
organoids were constructed and employed as a novel research model that could be used
instead of traditional cell lines and heterogeneous animal models.
Organoid
Organoid (Multiple Applications of Organoids)
The end of Chapter 1