Ch.

24 Biotechnology
○1 Biotechnology

- Any technological application that uses organisms, biological systems or

processes to produce goods and services.
○2 Selective breeding

- To produce animals or plants with desirable characteristics by selecting

the best individuals to breed.
Takes some time to produce the product with desired characteristics.
May not always work well
(i.e. the desired characteristics cannot be produced by this method.)
(Source: BioNinja)

○3 Genetic engineering- Recombinant DNA technology

- Contain DNA from different species.

- Involve techniques of recombinant DNA molecules construction:
- A gene (DNA fragment) is extracted and inserted to another DNA
molecule, mostly bacterial plasmid.
Δ Plasmid
- Small circular pieces of DNA separated from main bacterial
chromosome.
- Contains just a few genes.
- Help bacterium to overcome stressful situations (e.g. antibiotic resistant).
- Help the host to digest unusual substances or kill other types of bacteria.
- Replicate independently of chromosomal DNA.

Bacterial chromosome Plasmid

(Source: Wikipedia)

(Source: Kezia’s)
Δ Recombinant DNA Technology
Obtain DNA fragments that contain target gene
(e.g. gene coding for particular protein)
↓
1 2
Cut DNA fragments and plasmids with the same restriction enzyme
Restriction enzyme recognize specific base sequences
Cut DNA at specific points (i.e. same locus)
Cut open plasmids
↓
Insert target gene into plasmid with the help of DNA ligase (enzyme)
↓
Two pieces of DNA joint (ligation), producing recombinant plasmid (DNA)

Restriction Enzyme ≈ Scissor DNA Ligase ≈ Glue

○4 Gene cloning

- Host cell: cell for 1replication and 2expression of target gene.

(Usually use fast-growing microorganisms, e.g. E. coli and yeast)
- Vector: act as a vehicle to transfer genes, in this case, it is plasmid.
Recombinant plasmid taken up by host cell, replicates to produce identical copies
(including the target gene that it carries)
↓
Host cells divide, DNA molecules passed to daughter cells
Further plasmid replication takes place
↓
Clone of identical host cells produced
Each contains one or more copies of recombinant DNA molecules.

Restriction enzyme
+ DNA ligase
Plasmid Target gene Recombinant plasmid

Host cell

Replicate independently

Host cell divide, plasmid passed to daughter cells

(Source: Department of Bioinformatics, OUAT)

Note that
Plasmid can replicate independently. They are small and can move from one
bacterial cell to another easily. Hence, it makes it a useful vector.
○5 Applications of Recombinant DNA Technology

- Organism receiving the foreign gene is genetically modified organism

(GMO), which is not considered as new species, despite different
phenotype.
- Acquire new characteristics when foreign gene expressed through the
process of protein synthesis.
Medicine
- Genetically modified bacteria produce pharmaceutical products.
Δ Production of human insulin (treating diabetes):
Identify and introduce the gene that codes for human insulin to bacteria
by recombinant plasmid
↓
Host bacteria produce human insulin
DNA is universal among all organisms
↓
Large scale GM bacteria cultured in industrial fermentations
to produce large amount of human insulin

- Other pharmaceutical products:

Pharmaceutical products: Potential use:
Promoting growth in children with growth hormone
Human growth hormone
deficiency.
Blood clotting factor VIII Treating haemophilia.
Vaccines Treating hepatitis B, herpes, rabies and cholera
Interferons (干擾素) Treating cancer

- Used in gene therapy (treating genetic disorders by introducing normal

gene to cells with defective gene).
1 Cells with defective gene
□ 2 Virus is altered so it cannot
□
removed from patient. reproduce.

7 Genetically altered
□
cells carry out normal 3 Normal gene inserted
□
functions. into virus.
6 Altered cells injected
□
into patient.

4 Virus infect cells, inducing normal

□
gene to replace defective gene.

□5 Cells from patient

become genetically altered. (Source: biologyexams4u)

Cleaning up oil spills
- GM bacteria can break up hydrocarbons in oil.

(Source: )

Mining industry
- GM bacteria extract metals from their ores more efficiently than
conventional mining technologies.
Agriculture and food production
- GM crops: GM maize (herbicide resistance).
GM tomato (longer shelf life).
GM rice (higher content of carotene, a precursor of vitamin A)
- GM livestock: GM pigs
(produce omega-3 fatty acids, by placing roundworm gene to pigs)
GM salmon
(Higher growth rate)

(Source: Mike Keefe)

Note that
It is often danger to release GM organisms to the wild. They may out-compete wild
organisms or intensify the competition for resources.
For example, GM salmon have high growth rate and eats a lot. They may harm the
balance of ecosystem if released.
○6 DNA fingerprinting

- Technique used to identify individuals.

Δ Principle of DNA printing
- In our DNA, there are highly variable regions with repetitive base
sequence that do not contribute to the function of a gene.
- The length of these regions varies between individuals (except twins).
Thus, it can be used to identify individuals.
Δ Gel electrophoresis
Load samples of DNA fragments into wells of the gel
↓
Apply a voltage to drive DNA fragments across the gel
↓
DNA fragments move different distances according to their size

(Source: wikiversity)

- DNA fragments are negatively charged due to phosphate groups.

- Negatively charged (Cathode): DNA fragments placed
Positively charged (Anode): DNA moves towards
- Larger DNA fragments have less displacement than smaller DNA
fragments in the same period.
Note that
Gel slows down the motion of DNA fragments, so the pattern becomes observable.
Note that
Voltage applied cannot be too high, or else the blending would blur.

- After electrophoresis is completed, DNA bands can be visualized by

staining them with special dyes (e.g. fluorescent dye ethidium bromide
turn fluorescent under UV light when bonded to DNA).
- DNA fragments can also be labelled with radioisotope, DNA bands can
be copied to an X-ray film.
7 Applications of DNA Fingerprinting
○
Forensic science
- DNA in suspect’s blood/ other body material compared to that of the
evidence (e.g. hair, skin cells, semen).
- Prove guilt/ innocence in criminal trials.
Extract DNA from the sample (e.g. hair, skin cells, semen)
↓
Obtains DNA fragments (highly variable regions) by using restriction enzymes
↓
Separate DNA fragments by gel electrophoresis according to their molecular size
↓
DNA fragments can be revealed by staining
↓
Compare the DNA fingerprint formed
Prove guilt/ innocence of suspect

Parentage test
- Children receives half of genetic material from father and the other half
from mother. Mother Child Father

Victim identification
- Identifying the body of a victim (e.g after natural disasters).
- Applicable on badly decomposed body or a part of the body.
Evolutionary studies
- Assumed that closer evolutionary relationship of two groups of
organisms share more common places in their DNA fingerprints.
(Source: Pixabay)
○8 Human genome project (1990)

- Genome: All the genetic material of any cell in an organism, including

all genes and non-coding sections on all of the chromosomes.
- Complete knowledge of genome revolutionizes our understanding of life.
- Does not only study human genome, but also non-human organisms
(e.g. yeast, mouse, fruit fly)
Δ Goals of the project
- Determine DNA sequencing of around 3 billion bases of human genes.
- Identify and locate all around 20,000 human genes (gene mapping).
- Store genetic information in database and improve tools for data analysis.
- Transfer related technologies to private sector.
- Address the ethical, legal and social issues that arose from the project.
Δ Potential benefits
- Help scientist locate and identify genes related to genetic diseases
(e.g. heart diseases, cancer, Alzheimer’s disease).
- Enable scientists to assess health damage and risks caused by exposure to
mutagens and toxic agents (i.e. risk assessment).
- Develop technology that make use of microorganisms to produce energy,
toxic waste reduction, industrial processing (i.e. energy and industrial).
- Enable scientists to produce livestock and crops with improved
productivity and quality.
- Enhance evolutionary studies.
Δ Limitations Knowing sequences and location of genes

Knowing all biological processes in body

Knowing the role of every gene
Knowing the interactions with other genes
Knowing how DNA relates to behavior & brain function

Δ Concerns
- Ownership of personal genetic information (who can access?).
- Privacy of genetic information (persons who can access?).
- Psychological impact and public perception of individuals with genetic
differences, may cause discrimination.
(The way that genetic information affects society and individuals)

(Source: The Economist)