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27 Nutrition
Biotechnology
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Biotechnology
Think about…
27.1 Recombinant DNA technology
27.2 DNA fingerprinting
27.3 Human Genome Project
Recall Think about…
Concept map
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Supersweet maize Supersweet maize
No. In fact, a mutation

occurred naturally in
the maize plant.
This maize is so sweet!

grains store
Has the genetic make-up less starch and
of the maize plant been more sugars
modified by scientists?
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How can scientists modify the What kinds of genetically modified

genetic make-up of organisms? crops have been produced?
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Biotechnology (生物工程) Biotechnology (生物工程)

• refers to the use of organisms, • examples:
biological systems or processes to
produce goods or provide services
production of cheese production of wine

• now involves the use of recombinant
DNA technology (重組DNA技術)
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27.1 Recombinant DNA technology 6
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27.1 Recombinant DNA technology 27.1 Recombinant DNA technology

A What is recombinant DNA A What is recombinant DNA
technology? technology?
After an apple is cut…
an enzyme in apple tissue Now an apple that
catalyses the reaction does not turn brown
between oxygen and certain is available.
compounds in the tissue How is it possible?
à turns brown quickly
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27.1 Recombinant DNA technology • production of GMOs is an application

A What is recombinant DNA of genetic engineering (遺傳工程)
technology? changing of the genetic make-up of
an organism by direct manipulation
a gene was introduced into of DNA
the apple plant to stop its fruit
from producing the enzyme
recombinant DNA technology
that causes browning
is often used by scientists to
genetically modified
organism (GMO, introduce a new characteristic
ordinary apple GM apple 基因改造生物) into an organism
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B How does recombinant 1 Obtain DNA fragments containing

DNA technology work? Animation 27.1 the gene of interest
Recombinant DNA technology involves donor cell
4 major steps:
DNA containing gene of interest

gene of interest
(目標基因)
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2 Obtain vectors (載體) 2 Obtain vectors

a carrier to transfer the gene
of interest into a host cell for bacterium
expression
bacterial plasmid (質粒 )

chromosome a small ring of
extrachromosomal DNA
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3 Cut DNA fragments and plasmids 4 Join DNA fragments and plasmids
with a restriction enzyme (restriction) together using a DNA ligase (ligation)
restriction enzymes
gene of interest plasmid

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4 Join DNA fragments and plasmids

together using a DNA ligase (ligation)
introduced into
host cells for:
• producing GMOs with new characteristics,
e.g. the apple plants which produce fruits
recombinant plasmid recombinant plasmid
that do not turn brown when they are cut
(重組質粒 ) (重組質粒 )
• producing proteins of other species,
e.g. producing human insulin by bacteria
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C Applications of recombinant 1 Medicine

DNA technology a) Production of pharmaceutical products
Areas of application: e.g. human insulin
1 Medicine
• to control the blood glucose
2 Agriculture and food production level in diabetics
3 Industrial uses
4 Environmental protection
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Production of insulin Production of insulin

in the past in the past
• extracted from the pancreas of cattle or • problems:
sheep - takes a long time and at a high
extraction cost
- small amounts obtained from each
extraction
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in the past in the past
• problems: • problems:
- insulin from cattle or sheep has an amino - patients were at risk of infections if the
acid sequence slightly different from that cattle or sheep were infected by
of human insulin pathogens
à may be rejected by the immune system
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today today
• produced using recombinant DNA • by introducing the DNA encoding
technology human insulin into
bacteria
GM bacteria
cultured on a
large scale in fermenter (發酵器 )
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today today
extract polypeptides • advantages:
produced - takes a shorter time and at a lower
process into extraction cost
- much higher product yield
functional
human insulin
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Production of insulin Other pharmaceutical products

today produced using recombinant DNA
• advantages:
technology
- produces insulin that is structurally the Pharmaceutical
Use
product
same as the insulin produced by human
Human growth Treatment for growth hormone
body hormone deficiency
à not rejected by the immune system Hepatitis B vaccine Prevention of hepatitis (肝炎)
- produces pure insulin due to viral infection
à lower risk of causing infections
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Other pharmaceutical products 1 Medicine

produced using recombinant DNA b) Gene therapy (基因治療)
technology
• potential cure for diseases caused by
Pharmaceutical
product
Use a defective gene
Human blood Treatment for haemophilia
clotting factor VIII
(凝血因子VIII)
Human interferons Treatment for cancer
(干擾素)
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1 Medicine 2 Agriculture and food production

b) Gene therapy • to introduce desirable characteristics
vector into crops and farm animals
normal gene Purposes:

à supplement the • increasing the productivity of crops
defective gene and animals
cell with a • improving the nutritional value of
defective gene the foods
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2 Agriculture and food production 2 Agriculture and food production

• increasing the productivity • increasing the productivity
pest-resistant
GM maize plant
ordinary fast-
salmon growing
GM salmon
• grows to a marketable size in a • produces a pest-specific toxin
shorter time to kill certain pests
à productivity of fish farming é à crop loss due to the pests ê
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• increasing the productivity • increasing the productivity
herbicide-resistant
virus- GM soya bean plant
ordinary resistant
papaya GM papaya
(infected (not infected
by virus) by virus)
à crop loss due to
viral infection ê à crop loss due to the use of herbicides ê
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• improving the nutritional value • improving the nutritional value
- GM rice, rich in
β-carotene
converted to
vitamin A
Golden rice in our body Golden rice
ordinary rice
(黃金稻米 ) (黃金稻米 )
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• improving the nutritional value • improving the nutritional value
ordinary ordinary
pig pig
GM pigs GM pigs
• levels of unsaturated à eating their meat
fatty acids é may help prevent
heart disease
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2 Agriculture and food production Selective breeding

• Traditionally, crops and farm animals select organisms
with desirable characteristics are with the desirable
obtained by selective breeding characteristic
(選擇育種).
breed
apple plant apple plant
repeat
many times
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Selective breeding Selective breeding
After many
generations…
alleles for the desirable

characteristic become more and eventually all organisms in the population
more widespread in the population show the desirable characteristic
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In the past: Today:

• Nowadays, crops and farm animals with obtained by selective produced using
breeding recombinant DNA
desirable characteristics can be technology
produced using recombinant DNA • slower • quicker
technology. • involves many genes • involves only one or
Why? at the same time several genes at the
à results less same time
predictable à results more
predictable
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In the past: Today: 3 Industrial uses

obtained by selective produced using • to produce enzymes
breeding recombinant DNA
technology bacterium gene for enzyme
• cannot transfer genes • can transfer genes
between unrelated between unrelated
species species
produces
extracted
for use
enzyme
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3 Industrial uses 3 Industrial uses

• to produce enzymes e.g. • to produce enzymes
- digestive enzymes in biological GM bacterium
washing powders
- proteases in meat tenderizers
- enzymes used in cheese production produces • larger amounts
produced in a
- enzymes used in beer production
shorter time and
enzyme at a lower cost
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4 Environmental protection
• to help clean up oil spills
- GM oil-eating bacteria can break
1 Recombinant DNA technology:
down a wider range of hydrocarbons
• a technique in which a fragment
of DNA from a donor cell or
oil spills organism is isolated and then
inserted into the DNA of another
cell or organism.
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1 Recombinant DNA technology: 2 Major steps in recombinant DNA

• allows scientists to introduce a technology:
new characteristic into an a Obtain DNA fragments
organism by inserting a new containing the gene of interest
gene into its DNA. from donor cells.
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2 Major steps in recombinant DNA 2 Major steps in recombinant DNA

technology: technology:
b Obtain vectors (e.g. plasmids). c Cut the DNA fragments and the
plasmids using the same
restriction enzyme .
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2 Major steps in recombinant DNA 3 Applications of recombinant DNA

d Join the DNA fragments and the • To produce pharmaceutical
open plasmids together using a products and for gene therapy .
DNA ligase .
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3 Applications of recombinant DNA 3 Applications of recombinant DNA

• To increase productivity of • To produce GM bacteria that
crops and farm animals, and produce enzymes for industrial
improve the nutritional value uses.
of the foods.
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3 Applications of recombinant DNA
technology:
• To produce GM bacteria that
help to clean up oil spills .
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27.2 DNA fingerprinting 6
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A How does DNA fingerprinting

• 99.9% of the base sequence in DNA is the
work? called highly variable regions
same in all humans
• the remaining 0.1% is unique • there are many regions that contain repetitive
base sequences in our DNA
DNA fingerprinting
• between individuals:
(DNA 指紋分析 )
number of repeats in each region is different
individuals can be identified ® thus, lengths of these regions are different
analyzed by DNA fingerprinting

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A How does DNA fingerprinting 1 Extract DNA

work?
• involves 4 major steps:
DNA
blood sample
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2 Obtain DNA fragments 3 Separate DNA fragments
DNA fragments
separated according to
DNA fragments their size
containing different
highly variable regions gel electrophoresis
(凝膠電泳 )
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4 Produce DNA fingerprints Working principle of gel electrophoresis

• makes use of an electric field to drive
DNA fragments (negatively-charged)
DNA bands across a gel slab
• each band consists
of DNA fragments
of the same size
• unique to each
individual
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Working principle of gel electrophoresis Working principle of gel electrophoresis

negatively-
Animation 27.2 charged direction of movement
samples of DNA samples of DNA
fragments fragments
gel slab
– +
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with pores • DNA fragments are separated into
à shorter DNA fragments move faster bands according to their size
longer fragments shorter fragments longer fragments shorter fragments

(travel further) (travel further)
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• staining usually with fluorescent dye • the pattern of bands can be copied to a
allows the bands photographic film
to become visible
bands become
visible under
UV light
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27.1 27.1
Practical 27.1
Separation of DNA fragments Procedure

using gel electrophoresis A Preparation of 1X TBE buffer solution
glass rod to mix well
80 cm 3 5X TBE buffer
solution + 320 cm 3
gel electrophoresis kit distilled water
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27.1 27.1
B Preparation of agarose gel 2 Heat for one minute until the agarose
powder completely dissolves.
1
swirl to mix well microwave
oven
60 cm 3 1X TBE buffer
solution + 0.9 g agarose
powder
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27.1 27.1
3 Cool down the solution to about 60 °C. 4
sticky tape gel mould
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27.1 27.1
4 5 gel comb
agarose
solution
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27.1 27.1
5 6 Remove the gel comb and sticky tape after
30 minutes.
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27.1 27.1
C Gel electrophoresis of DNA fragments 2
1 Label the six tubes of DNA samples A to F. gel electrophoresis tank
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27.1 27.1
3 4 syringe (or micropipette)
1X TBE buffer solution
fitted with a tip
DNA
marker right
left
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27.1 27.1
4 syringe (or micropipette) 4 Wash the syringe with a new buffer solution.
fitted with a tip
Load DNA samples A to F into separate wells.
DNA
marker right Wash the syringe with a new buffer solution
several times between successive loadings.
contains DNA fragments of known sizes
à used to estimate the sizes of Change a new tip if a micropipette is used.
DNA fragments
left
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27.1 27.1
4 right 5 100 V d.c.
– +
DNA samples
A to F lid
DNA marker left

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27.1 27.1
100 supply.
6 Turn off the power V d.c. 7 Wait for about 12 hours.
tracking dyes methylene blue
solution
– +
gel
~2/3 of gel length plastic box

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27.1 B Applications of DNA

8 Wash the gel with running water and fingerprinting
take a photograph of it.
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1 Forensic science 1 Forensic science

• to provide evidence of the identities of DNA fingerprints from a murder case:
individuals in court cases
sample from
blood sample victim crime scene suspect 1 suspect 2 suspect 3
collected from
crime scene
produce a DNA
fingerprint
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1 Forensic science 1 Forensic science

DNA fingerprints from a murder case: DNA fingerprints from a murder case:
sample from sample from

victim crime scene suspect 1 suspect 2 suspect 3 victim crime scene suspect 1 suspect 2 suspect 3
sample Note:
collected from • the presence of DNA in the crime scene
crime scene
does not prove that suspect 3 is the murderer
likely to have
come from • the DNA fingerprints should be considered
suspect 3 along with other types of evidence
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2 Parentage tests 2 Parentage tests

• to establish family relationship Animation 27.3
• to establish family relationship
father mother child 1 child 2 child 3 father mother child 1 child 2 child 3
half half
bands bands
from from
mother father
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3 Victim identification 4 Authentication of foods and

• to compare the DNA fingerprint of the dead Chinese medicines
body with that of the suspected victim • to identify substituted (冒充的 ) Chinese
medicines by comparing
the DNA fingerprint
of an unknown sample
with a genuine one
substitutes of cordycep
(蟲草 ) are commonly found
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4 Authentication of foods and 5 Conservation of endangered

Chinese medicines species
• to identify the types of meat in a meat • to trace the origin
product by producing a DNA fingerprint of of ivory products
a meat product and and help combat
comparing the illegal hunting of
band pattern with African elephants
those of different
types of meat
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6 Tracing the source of infectious 7 Screening for genetic diseases

diseases • to help diagnosis of genetic diseases in
• to quickly identify the strains of bacteria prenatal and new born babies
that cause certain infectious diseases
by comparing DNA fingerprints of prenatal
and new born babies with those derived from
help trace where the bacteria have come from
the normal allele and the diseased allele of
the gene associated with the genetic disease
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6 Nutrition in humans 6 Nutrition in humans
In a parentage test, the DNA fingerprints of a boy

(X), his mother and a man (P) were prepared. boy (X) mother man P girl (Y)
The diagram on the next slide shows their DNA
fingerprints. The DNA fingerprint of a girl (Y),
a sister of X, is also included.
115 116
a Explain why DNA fingerprinting can be used in Suggested answer

parentage tests. (1 mark) a Half the genetic material of an individual comes
from the father and the other half from the mother.
(1)
117 118
b Based on the DNA fingerprints, deduce whether Suggested answer

P is the biological father of the boy. Explain your b He is not the biological father of the boy. (1)
answer. (3 marks)
Some of the bands in the boy’s DNA fingerprint
boy (X) mother man P girl (Y) resemble the pattern of his mother’s DNA
fingerprint, (1)
while the other bands cannot be found in P’s DNA
fingerprint. (1)
119 120
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Interpreting DNA fingerprints Interpreting DNA fingerprints

boy (X) mother man P girl (Y) Step 1 boy (X) mother man P girl (Y)
Is he the align the highlight

biological bands of the bands
father of X? the boy in common
with those
of his
mother
121 122
Interpreting DNA fingerprints Interpreting DNA fingerprints

Step 2 boy (X) mother man P girl (Y) boy (X) mother man P girl (Y)
Is he the
biological
compare father of X?
the no match no match Not likely.
remaining
bands with
those of P no match no match
123 124
The diagram below shows the DNA fingerprints of State which of the children is likely to be
a couple and three children.
a the biological child of the couple. (1 mark)
Child 3 (1)
husband wife child 1 child 2 child 3 husband wife child 1 child 2 child 3
125 126
21
State which of the children is likely to be State which of the children is likely to be
b a child from the wife's previous marriage. (1 mark) c a child unrelated to the couple. (1 mark)
Child 2 (1) Child 1 (1)
husband wife child 1 child 2 child 3 husband wife child 1 child 2 child 3
127 128
c i Give one evidence to support that Y is the Suggested answer

biological sister of X. (1 mark) c i Half of the bands in Y’s DNA fingerprint
resemble the pattern in the DNA fingerprint of
X’s mother. (1)
boy (X) mother man P girl (Y)
129 130
c ii Based on the events that happen during Suggested answer

gamete formation and fertilization, explain c ii Due to independent assortment / crossing over
why the DNA fingerprints of X and Y are of chromosomes, (1)
different. (3 marks)
boy (X) mother man P girl (Y) gametes produced from the same person are
genetically different. (1)
Random fertilization of gametes results in
individuals of different genetic composition. (1)
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1 DNA fingerprinting is a 2 Major steps in DNA fingerprinting:

technique to identify an individual a Extract DNA from samples.
based on a DNA sample. b Obtain DNA fragments
containing the selected regions
from the DNA sample.
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2 Major steps in DNA fingerprinting: 3 Gel electrophoresis makes use of

c Separate DNA fragments using an electric field to drive DNA
gel electrophoresis . fragments across a gel slab
d Produce DNA fingerprints . towards the positive terminal.
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3 The DNA fragments are finally 4 DNA fingerprinting can be used:

separated according to their • in forensic science to provide
size . evidence of the identities of
individuals in court cases.
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4 DNA fingerprinting can be used: 4 DNA fingerprinting can be used:

• to establish family relationships • to identify victims in disasters.
in parentage tests .
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• to authenticate foods and • to help conserve endangered
Chinese medicines. species , e.g. by tracing the
origin of ivory products.
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• to trace the source of • to screen for genetic diseases.
infectious diseases.
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27.3 Human Genome Project 6
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27.3 Human Genome Project A What is the HGP?

(HGP) the complete set of • started in 1990
DNA in humans • involved scientists from 18 countries
with advances in
DNA sequencing
technologies
the base sequence of it

can be determined
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A What is the HGP? A What is the HGP?

• goals: • goals:
- to determine the base sequence of - to store the information in an open
human genome database
- to locate all the genes (gene mapping - to develop related new technologies and
定位 ) improve tools for data analysis
- to address the ethical (道德倫理的 ),
legal and social issues
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1990 1996
HGP started in the US sequencing of yeast and
mapping of mouse genome
completed
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1999 2000
sequencing of first human draft version of human
chromosome completed genome sequence
completed
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2003 all goals achieved,

finished version of human but analyses of
genome sequence completed data continue
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Found out that:
• there are ~ 3.2 billion base pairs in the
human genome.
• there are ~ 21 000 genes identified in the
human genome.
printout of the human genome

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A What is the HGP? B Limitations of the HGP

Found out that: • functions of many genes are still
• only ~ 1.5% of our DNA codes for proteins. unknown
The other 98.5% is non-coding DNA.
• ~ 99.9% of the DNA is almost exactly the • data obtained are still not enough to
same in all people. understand some biological processes,
e.g. brain function
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C Benefits of the HGP C Benefits of the HGP

1 Better understanding of genetics 3 Better understanding of evolution
2 Improved diagnoses and treatment of
diseases
e.g. breast cancer and Alzheimer’s
disease
98.6% 85%
similarity in base sequence
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C Benefits of the HGP

3 Better understanding of evolution 1 Goals of the HGP:
• To sequence the human
genome.
• To map all the genes in the
human genome.
more closely related
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1 Goals of the HGP: 1 Goals of the HGP:

• To store the information in a • To address the ethical , legal
database. and social issues that may arise.
• To develop related new
technologies and improve
tools for data analysis.
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2 Benefits of the HGP: 2 Benefits of the HGP:

• Better understanding of • Better understanding of
genetics . evolution .
• Improved diagnoses and
treatment of diseases.
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1 How can scientists modify the genetic

2 Limitations of the HGP: make-up of organisms?
• The functions of many genes
Scientists can use recombinant DNA
are still unknown. technology to introduce DNA into
• The data obtained is still not organisms.
enough to understand some
processes.
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2 What kinds of genetically modified crops Biotechnology

have been produced?
some developments
Virus-resistant papaya plants, herbicide- include
resistant cotton plants, etc.
recombinant Human
DNA
DNA Genome
fingerprinting
technology Project (HGP)
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recombinant DNA DNA fingerprinting

technology applications include
applied in
forensic victim
production of science identification
genetically modified
organisms (GMOs) parentage authentication of foods
tests and Chinese medicines
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DNA fingerprinting Human Genome

applications include Project (HGP)
data contributes to
conservation of screen for
endangered genetic better understanding better understanding
species diseases of genetics of evolution
trace the source of improved diagnoses and

infectious diseases treatment of diseases
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27

Supersweet maize Supersweet maize

No. In fact, a mutation

This maize is so sweet!

How can scientists modify the What kinds of genetically modified

Biotechnology (生物工程) Biotechnology (生物工程)

production of cheese production of wine

27.1 Recombinant DNA technology 27.1 Recombinant DNA technology

27.1 Recombinant DNA technology • production of GMOs is an application

B How does recombinant 1 Obtain DNA fragments containing

DNA containing gene of interest

2 Obtain vectors (載體) 2 Obtain vectors

bacterial plasmid (質粒 )

gene of interest plasmid

4 Join DNA fragments and plasmids

C Applications of recombinant 1 Medicine

Production of insulin Production of insulin

Production of insulin Production of insulin

Production of insulin Production of insulin

Production of insulin Production of insulin

Production of insulin Other pharmaceutical products

Other pharmaceutical products 1 Medicine

1 Medicine 2 Agriculture and food production

normal gene Purposes:

2 Agriculture and food production 2 Agriculture and food production

2 Agriculture and food production 2 Agriculture and food production

2 Agriculture and food production 2 Agriculture and food production

2 Agriculture and food production 2 Agriculture and food production

2 Agriculture and food production Selective breeding

Selective breeding Selective breeding

alleles for the desirable

In the past: Today:

In the past: Today: 3 Industrial uses

3 Industrial uses 3 Industrial uses

1 Recombinant DNA technology: 2 Major steps in recombinant DNA

2 Major steps in recombinant DNA 2 Major steps in recombinant DNA

2 Major steps in recombinant DNA 3 Applications of recombinant DNA

3 Applications of recombinant DNA 3 Applications of recombinant DNA

27.2 DNA fingerprinting

A How does DNA fingerprinting

analyzed by DNA fingerprinting

A How does DNA fingerprinting 1 Extract DNA

2 Obtain DNA fragments 3 Separate DNA fragments

4 Produce DNA fingerprints Working principle of gel electrophoresis

Working principle of gel electrophoresis Working principle of gel electrophoresis

Working principle of gel electrophoresis Working principle of gel electrophoresis

longer fragments shorter fragments longer fragments shorter fragments

Working principle of gel electrophoresis Working principle of gel electrophoresis

Separation of DNA fragments Procedure

DNA marker left

~2/3 of gel length plastic box

27.1 B Applications of DNA

1 Forensic science 1 Forensic science

1 Forensic science 1 Forensic science

sample from sample from

2 Parentage tests 2 Parentage tests

3 Victim identification 4 Authentication of foods and

4 Authentication of foods and 5 Conservation of endangered

6 Tracing the source of infectious 7 Screening for genetic diseases

In a parentage test, the DNA fingerprints of a boy

6 Nutrition in humans 6 Nutrition in humans

a Explain why DNA fingerprinting can be used in Suggested answer

6 Nutrition in humans 6 Nutrition in humans