Genetics The Code Broken?

(read these alongside Blueprint of Life notes)

The structure of a gene provides the code for a polypeptide
1. Describe the processes involved in the transfer of information from DNA
through RNA to the production of a sequence of amino acids in a polypeptide
---------------------See BLUEPRINT OF LIFE notes, Focus 4, Point 2-----------------------2. Choose equipment or resources to perform a first-hand investigation to
construct a model of DNA
DNA is a double-stranded molecule twisted into a helix with each strand comprised
of a sugar-phosphate backbone and attached bases adenine (A), thymine (T),
cytosine (C) and guanine (G) connected to a complementary strand by pairing the
bases, A-T and G-C
---------------------------------Constructed DNA Models in
Class---------------------------------PRAC EXPERIMENT 1!
3. Process information from secondary data to outline the current understanding
of gene expression
A gene is a piece of a DNA molecule that codes for a specific polypeptide. The
sequence of bases on the portion of the DNA molecule (gene) ultimately determines
the sequence of amino acids in polypeptide produced. These join to form proteins
such as enzymes, hormones and structural proteins in cell membranes. Proteins
control all chemical reactions and hence all cell activities. In this way, genes
determine the characteristics of an organism. Gene expression occurs through
polypeptide synthesis, protein synthesis and the functioning of that protein.

Gene expression: The conversion of the information on a gene to a protein

product resulting in a cell characteristic.
Gene expression refers to a gene being activated to produce a polypeptide.
They can be switched on and off by various mechanisms.
In multicellular organisms, differentiated cells only have a small number of their
genes activated. E.g. muscle cells only have those genes activated that control
muscle factors.
Some genes are active for defined periods of development while others are active
throughout all stages of development of an organism.
Some genes are active in cells of specific tissues only while other genes are
active in all cells.
For this to have occurred, gene expression must have been regulated.

Gene expression must be controlled:

- To prevent mutation
- To complete in the correct sequence
- To make the correct DNA
The Process of Gene Expression:
Most of the control of the activation and deactivation of genes occurs in the
transcription of DNA into RNA. However control must be ministered during the
activation and deactivation of the chromosome (unwinding the DNA molecule), RNA
transport, initiation of translation and protein activity.
a) DNA is a tightly wound molecule so that large amounts of information can be
stored in a small area. The parts that are very highly condensed are the genes
that are switched off, sometimes even permanently switched off.
b) Promoters are a regulatory area at the start of a gene that acts as the binding
site for RNA polymerase (makes mRNA) in unravelling DNA.
c) If there is a problem with the promoter there will be a problem in starting
transcription and thus the polypeptide will not be synthesised and the gene will
not be expressed. If the binding strength of the promoter region is increased,
there will be a greater quantity of the protein produced.
d) The presence of some chemical groups can block gene expression. Methyl
groups block DNA from expression and acetyl groups allow gene expression to
occur more easily.
e) A large amount of DNA (and therefore mRNA) is not coded into protein. The noncoding nucleotides (introns) are interspersed between the coding nucleotides
(exons). The introns must be cut out before the mRNA leaves the nucleus, then
the exons spliced together. Regulatory proteins control the splicing and may be
different in different cells.
f) Ends of mRNA are capped with protective nucleotides that prevent breakdown
by hydrolytic enzymes and help mRNA to attach to the ribosome. Removal of
these nucleotides shortens the life of the mRNA so the protein synthesis is
stopped.
g) Translation of the mRNA can be blocked if specific proteins bind to the mRNA in
the cytoplasm so that it cannot attach to the ribosome

Multiple alleles and polygenic inheritance provide further variability within a

trait
1. Give examples of characteristics determined by multiple alleles in an organism
other than humans
Alleles are variations of a gene which are found at the same locus. A locus is a
particular place or position on a particular chromosome.
Sometimes more than 2 alleles exist in a population (multiple alleles) but each
individual has only 2 places or loci where these genes are located on a part of
homologous chromosomes. Where there are multiple alleles, the number of
variations in a population is greater.

Examples:
White clover leaf patterns there are 7 alleles for the patterns on the leaves. This
gives 22 possible phenotypes
White markings on cattle there are 4 alleles and 10 possible genotypes
In general, the more number of alleles that exist for a gene allow for a greater the number of
possible genotypes and phenotypes and therefore greater amount of variation within population.
2. Compare the inheritance of the ABO and Rhesus blood groups
There are three alleles for the human blood. A and B are co-dominant, O is
recessive. Any individual can hold any of these three alleles but can only carry two of
these three alleles.
AA or IAIA is phenotype A
AB or IAIB is phenotype AB
BB or IBIB is phenotype B

AO or IAi is phenotype A
BO or IBi is phenotype B
OO or ii is phenotype O

There are 3 alleles that determine human blood, A, B and O.

There are 6 possible genotypes AA, AB, BB, AO, BO, OO.
There are four possible phenotypes A, B, AB, O.

Two antigen and two antibodies are mostly responsible for the ABO types. The
antigens are on the surface of the red blood cells and the antibodies are in the blood
plasma.
A
B
AB
O

A blood
carries the
antibodies for
B blood

B blood
carries the
antibodies for
A blood

Fights B and
AB blood

Fights A and
AB blood

AB blood
carries no
antibodies

O blood
carries
antibodies for
A and B
blood

Accepts all blood

Fights A, B
types however
and AB blood
cannot donate to
only accepts
any other blood
O blood.
type.
If red blood cells carrying one or both antigens contact their corresponding
antibodies e.g. antigen A contacts Anti-A, the red blood cells agglutinate (clump
together) which causes death.
O blood is the universal donor and so it can be used all other blood types.
AB blood is the universal receiver as it can receive blood from all other groups.
Rhesus Factor
-

This is a group of transmembrane antigens found on the surface of red blood

cells.
You are either Rh positive (Rh+) meaning that you have the factor or Rh negative
(Rh) meaning that you do not have the factor but produce antibodies if your
blood comes in contact with this factor.
This is completely separate from the ABO antigens, although it is found of red
blood cells. It is controlled by a different gene found in Chromosome 1. Rh + is
dominant over Rh.

Problems can arise if an Rh mother is carrying an Rh+ foetus. If small amounts of

foetal blood cross the placenta (usually in late pregnancy or at birth) antibodies can
be formed in the mothers blood against the foetus.
The danger does not occur for the first baby but in subsequent pregnancies when
the mothers blood will have memory cells producing antibodies against the Rh +
factor. These antibodies cross the placenta and start to attack and kill the red
blood cells of the foetus.
As a result, the child will be born with haemolytic disease of the newborn and will
have very few red blood cells and could die.
Only some types of antibodies can cross the placental membrane and some
cannot. The anti-Rh antibodies can cross the placenta. The anti-ABO antibodies
are different and cannot. In this way, if the mother and the child have different
blood types they will not be affected.
Inheritance of the Rh factor:
The Rh factor involves three different pairs of alleles which are located on three
different loci on chromosome pair number 1.
It is the combination of the 3 letters that determine the allele

In the diagram there are three pairs of Rhesus factor alleles, C and c, D and d, E
and e.
Possible genotypes will have either one C or c, D or d and one E or e from each
chromosome.
The outcome of predicting the Rhesus factor of an individual can be achieved by
using a trihybrid cross.
Genotype combinations with DD or Dd are classifies as Rh + and genotype
combinations with dd are classified as RhE.g.
Two parents each have CDE/cde and cdE/cDE in their germ cells. What is the
chance of them producing a child with Rh+ or Rh?

Compare results with a given table to determine whether the possible

characteristics are Rh+ or Rh.
Therefore there is a 25% chance that their child will be Rh and a 75% chance
that their child will be Rh+.

3. Define what is meant by polygenic inheritance and describe one example of

polygenic inheritance in humans or another organism.
Polygenic inheritance is where a particular trait is influenced by a number of
pairs of alleles which occupy different positions (loci) on chromosomes. The effect
of these genes are added together to produce a phenotype.
E.g. Human height is the result of a combination of genes
Discontinuous variation: result of several alleles on the same gene (or a small
number of genes) portraying the one trait.
Continuous variation: result of various alleles on different genes combining to
display a trait.
In any given population, the majority of individuals will not show the extremes
of the condition, but will be grouped together near the average in a normal
distribution or bell shaped curve when graphed.
This type of inheritance is also referred to as continuous variation and
displays evidence for polygenic inheritance.
Polygenic inheritance usually produces a large variation in phenotypes
Examples include:
Skin colour in humans
Length of corn cob
Size of hens eggs
Butterfat content of milk in cows
Colour of wheat kernels
Human Height:
Human height is controlled by perhaps ten or more genes
However appearances, especially body size is determined not only by genotype
but also by the environment e.g. poor nutrition may inhibit a person from
reaching their full height potential
If the height of men was surveyed and plotted, from height against number of
men with that height, the resultant graph would have a bell shaped curve
showing a normal distribution between the extremes of short and tall.
Environmental conditions often cause the shape of the graph to become a
smooth curve.
The greater number of genes involved, the greater the variation in the possible
phenotypes.
Plant Height in Tobacco:
This is controlled by a series of genes at multiple loci.
To quantitatively predict the appearance of offspring several assumptions are
made in the polygenic model of inheritance.
For example, it is assumed that each gene has a small but approximately equal
effect, the effects of each allele are additive, there is no linkage and the
environmental influences are ignored.

4. Outline the use of highly variable genes for DNA fingerprinting of forensic
samples, for paternity testing and for determining the pedigree of animals
DNA fingerprinting:
This is when scientists extract DNA from a sample of blood, skin, hair, semen
and other forensic samples.
They use restriction enzymes to cut DNA into fragments, which they separate by
a process called gel electrophoresis
Gel electrophoresis is where DNA is placed on a plate of gel and an electrical
current is applied.
Since DNA is negatively charged, it moves towards the positive electrode; the
smaller the fragment, the faster it moves and so the fragment separates
according to size.
Where they stop an ink blot is made on a piece of filter paper

The separated fragments are analysed by a process called southern blotting

which is where a sheet of nitrocellulose paper membrane is placed on the gel

and covered with filter paper.
The strands in each DNA segment are separated and a radioactively labelled
gene probe is added. Finally, the membrane is place on an X-ray film. The probe
makes an image on the film, showing where bands are located.
The result is called a genetic fingerprint.

DNA fingerprints cannot identify a person with absolute certainty, but the probability of
Pedigrees
animals:
identifying
thefor
correct
person is very high. However, there are arguments concerning this
DNA fingerprinting is also used byprobability
animal breeders to accurately identify
parentage of offspring.
This means that accurate records of pedigrees can be kept.
In some countries, dog breeders use DNA fingerprinting to guarantee bloodlines
for a particular breed
Paternity Testing:
No two humans have identical DNA, except for identical twins.
DNA fingerprinting or profiling can determine the parents of a child. DNA
fingerprinting relies of changes in sequences in DNA, known as polymorphisms.
In a paternity case, the following procedure is used:
1)
2)
3)
4)

DNA is collected from the nucleus

DNA is cut with restriction enzymes, which targets DNA polymorphisms
DNA fragments are separated using electrophoresis
The products of electrophoresis are transferred to a membrane by the Southern
blotting technique
5) The membrane from the blot is incubated with radioactive DNA, which binds to
specific sequences on the membrane
6) After chemical luminescence or autoradiography, the DNA, banding pattern or
fingerprint is revealed
7) The banding pattern of the mother, father and child are compared
If a parent has no bands in common with the child, then the parent could not be
the parent of this child.
If the parent shares a banding pattern and an allele with the child, it supports, but
does not prove that the parent is the parent of the child.
The error can be minimised with the greater number of different DNA probes. If the
data from the different probes result in the parent sharing more banding patterns and
alleles with the child, then it is more likely that the parent is the parent of that child.
5. Solve problems to predict the inheritance patterns of ABO blood groups and
the Rhesus factor
-----------------------------------------------See Point 2------------------------------------------------------------------------------------See Multiple Alleles Worksheet---------------------------------6. Process information from secondary sources to identify and describe one
example of polygenic inheritance
------------------------------------------------See Point 3-------------------------------------------------

Studies of offspring reflect the inheritance of genes on different chromosomes

and genes on the same chromosomes
1. Use the terms diploid and haploid to describe somatic and gametic cells

Diploid: Number of chromosomes possessed by an organisms body

cells/somatic cells (e.g. humans: 2n = 46)

Haploid: Number of chromosomes possessed by an organisms sex

cells/gametic cells normally half the number of chromosomes as in an
organisms body cells (e.g. humans: n = 23)

2. Describe outcomes of dihybrid crosses involving simple dominance using

Mendels explanations
---------------------------See Blueprint of Life Notes, Focus 2, Point 1-------------------------

Dependant Assortment: Alleles that are dependent on each other so are not
separated but instead transmitted to the gametes together. This shows that the
genes for an allele are on the same chromosome.

Independent Assortment: Alleles that are independent of each other so can be

separated and transmitted to the gametes independently. This shows that the
genes for an allele are on different chromosomes.

Mendels experiment of INDEPENDANT ASSORTMENT:

Mendel crossed a pure breeding round, yellow seeded plants with a pure
breeding wrinkled, green seeded plants.(Parental generation)
Round trait is dominant to wrinkled and yellow trait is dominant to green.
The offspring all produced round and yellow seeds. (F 1)
The F1 generation was crossed to obtain an F2 generation
The F2 generation contained totally new combination of plants that appeared in
the ratio of 9 yellow round:3 yellow wrinkled:3 green round:1 green wrinkled
(9:3:3:1)
This could only have occurred if the genes were allowed to combine at random
(an allele of a gene pair can combine with either allele of another gene pair)

Gametes

RY

Ry

rY

ry

RY

RRYY

RRYy

RrYY

RrYy

Ry

RRYy

RRyy

RrYy

Rryy

rY

RrYY

RrYr

rrYY

rrYy

Ry

RrYy

Rryy

rrYy

rryy

F2 genotypes:
or

rrYy or

RRYY or

RRYy or

RRyy or

RrYY or

RrYy or

Rryy or

rryy

F2 phenotypes:

round yellow
wrinkled yellow
round green
wrinkled

green
3. Predict the difference in
inheritance patterns if two
genes are linked
Linked: Two or more genes
are located close together
on the same chromosome
and are thus often inherited
together.
Linkage reduces the variety of
offspring that can be produced.

Phenotype ratio: 9:3:3:1

rrYY

Testing for linked genes:

A test cross is carried out between a double heterozygote (AaBb) with a
double homozygous recessive (aabb)
Unlinked:
If the two gene loci are not linked, the gene will assort independently and the
outcome of the test cross will be four classes of offspring in equal proportions.
ab
Ratio:

AB
AaBb
25%

ab
aabb
25%

Ab
Aabb
25%

aB
aaBb
25%

This means that the characteristic is unlinked so occurs on two separate

chromosomes.
Linked:
If the two loci are linked and are not separated by a great distance (less than 40 map
units), the outcome of the test cross can reveal the linkage.
The proportions will not be equal, instead there will be an excess of offspring from
parental type gametes and a deficiency of offspring from recombinant type gametes.
ab
Ratio:

AB
AaBb
40%

ab
aabb
40%

Ab
Aabb
10%

aB
aaBb
10%

This is because there is a higher chance of holding the parental characteristics and a
lesser chance of holding recombinant type gametes when genes are linked.
- But it depends on the heterozygous parent which allele genes are together on
one chromosome
From the proportion of recombinant offspring, the separation distance of the linked
genes on a chromosome (in map units or centimorgans/cM) can be estimated using
the formula:

These test crosses can be used to show if two genes are linked and how far apart
they are.

4. Explain how cross-breeding experiments can identify the relative position of

linked genes
------------------------------------------------See Point 3------------------------------------------------5. Discuss the role of chromosome mapping in identifying relationships between
species
Breeding experiments not only tell us which genes are on the same chromosome,
they can also tell us the exact order of these genes on the particular chromosome
and how close they are to each other.
This is done by studying the percentage of recombinant types among offspring.
If recombination occurs 5% of the time, then the two genes are said to be 5 units
apart on a chromosome. These are used to draw linkage maps.
This information also allows us to compare chromosomes from different species to
estimate their evolutionary distance from each other.
Once maps have been prepared for two species, computer programs can be used
to find the degree of similarity between them. The more similar the chromosome
maps, the closer related.
6. Process information from secondary sources to analyse the outcome of
dihybrid crosses when both traits are inherited independently and when they
are linked
------------------------------------------------See Point 3------------------------------------------------7. Perform a first-hand investigation to model linkage
PRAC EXPERIMENT 2!
The Human Genome Project is attempting to identify the position of genes on
chromosomes through whole genome sequencing
1. Discuss the benefits of the Human Genome Project
The aim of the Human Genome project was to map and understand the genes of
human beings.
The benefits of the Human Genome Project included allowing better understanding
into:
Treating human disease through molecular medicine
Providing information on the location of and identification of the genes that cause
diseases, such as inherited colon cancer, familial breast cancer and cystic
fibrosis, will lead to improvements in their treatment and prevention.

Assessing the risks of diseases

Genetic differences between people make some people more resilient to disease
so understanding more about this genetic variability will allow scientists to

estimate the health damage and risk of a disease that could occur from exposure
to low-level radiation, cancer-causing agents and inheritable mutations.
Determining evolutionary relationships between humans and other organisms
This allows a better understanding of evolutionary relationships, evolutionary
history similarities in diseases and immune systems between humans and other
organisms and similarities and differences between eukaryotes and prokaryotes.
Making medicines specific for an individual
Allow for development of preventative medicine tailored for the individual, health
risks for individuals and suggesting lifestyle changes which can lead to a
healthier life.

Identifying similarities and differences in DNA for forensic purposes

Can help determine between people responsible for crimes rather and those that
are innocent bystanders. Donors of organs will be better matched and paternity
of children will be easier to determine. The authenticity of food will also be clear.

Use bacterial genomes to provide new energy sources

Potential for bacterial genomics for use as biofuels, carbon recycling,
environmental remediation and in the understanding of pathogenic diseases.

Create disease-resistant plants and animals for agricultural production.

Producing insect resistant and disease resistant plants as well as animals that
are cheaper to produce and may have better nutritional value.

2. Describe and explain the limitations of data obtained from the Human Genome
Project
Limitations of the data obtained from the Human Genome:
There are many sequences of repetitive DNA and their purpose is not well
understood.
There are also ethical and social issues, including issues in privacy and
confidentiality and the use of genetic information.
Such as if a patients health history and genetic information is known by
insurance companies, they may be at the risk of higher premiums or may not be
provided with cover because of pre-existing conditions.
Impact of knowing as a child, you are likely to have a certain disease may
influence your attitude to life.
There is also debate about genetics and their interaction with the environment in
triggering diseases, a positive test result for a disease does not guarantee that it
will occur and vice versa.
3. Outline the procedure to produce recombinant DNA
1) A target gene is cut out of the DNA of one species through use of a restriction
enzyme
2) A plasmid is removed from a bacteria cell and is cut open using the same
restriction enzyme (therefore cutting at the same sequence)

3) Plasmid and target gene are combined in a test tube and the enzyme ligase
sticks the two together
4) Plasmid is inserted back into the bacterium
5) The bacterium reproduces, thus producing more copies of the target gene
6) The bacterium expresses the target gene.
-------------------------See Blueprint of Life Notes: Focus 5, Point 2---------------------------4. Explain how the use of recombinant DNA technology can identify the position
of a gene on a chromosome
Probe: A specific sequence of DNA that is complementary to the other DNA bases

on a gene or part of a gene.

a) A section of DNA with a known sequence of gene bases is cut (probe) and many
copies of the probe are made using recombinant DNA technology.
b) Fluorescent markers (dye) are attached to the probes
c) Chromosomes are treated to separate the strands
d) Probes are added to treated chromosomes
e) Probes hybridise (bind) to the DNA following the complementary base pairing rule.
A fluorescent light is shone on the hybridised DNA and the tagged probes which
are attached to a location on a chromosome glow brightly showing which
chromosome a particular gene is located. The position of that gene can be
observed and recorded.
\

This technique can be used to determine which chromosome the gene belongs to
and the specific position of the gene on the chromosome. This also helps scientists
in identify abnormal chromosomes and defective genes in a process called genetic
screening.
-----------------------------------------See Focus 3, Point
3-------------------------------------------5. Process information from secondary sources to assess the reasons why the
Human Genome Project could not be achieved by studying linkage maps
Linkage maps consist of genes that have been mapped and isolated according to
hereditary information.
They are determined by studying the chances of linked genes staying together
during crossing over and thus do not give an accurate position of the genes on a
chromosome.
The human genome consists of thousands of genes that are responsible for every
characteristic and function of the body and so linkage maps would have only
provided information about some clear hereditary factors for the Human Genome
Project and not for all the other many genes.
Linkage maps also do not account for the interaction of many genes for
expression of a phenotype.
Also as linkage maps also do not provide detailed base sequences and segments
of DNA vary largely due to overlaps and repeats, the
process of identifying the position of genes is too
complex to be achieved by studying linkage maps.
Gene therapy is possible once the genes responsible
for harmful conditions are identified
1. Describe current use of gene therapy for an
identified disease

Gene therapy: the transfer of a gene or genes into the

somatic cell of a patient to repair a genetic defect or to
introduce a novel function for the purpose of curing or
alleviating disease.

There are three approaches to gene therapy:

Gene addition a normal gene is inserted to

compensate for a non-functional gene

Gene compensation an abnormal gene is repaired

through selective reverse mutation (very tricky)

Gene knockdown the gene expression of an abnormal

gene is shutdown at mRNA level.
Gene therapy is only completed in somatic cells as these
genes are not passed onto the next generation. Gene therapy

on gametes is passed onto the next generation so for ethical reasons, this type of therapy
is not allowed in humans.
Process of Gene therapy:
1)
An abnormal gene is identified
2)
A normal copy of the gene is isolated
3)
The normal gene is encapsulated in a vector (e.g. a virus) that will carry it into the
target cell.
4)
The gene is delivered to the target cell. This may be done ex vivo (outside the body)
or in vivo (inside the body)
5) Scientists
must determine
if the
gene is being
.
Viruses
(retroviruses
or new
adenoviruses):
expressed (protein being made) in sufficient quantities to
.
Consist of DNA or RNA enclosed in a protein coat.
affect the phenotype.
They bind to cells then introduce their genetic material into the cell. The cell is
then forced to replicate the foreign virus DNA and express any proteins that are
coded for in the genetic material.
This makes them ideal vectors to deliver foreign genes into a cell.
Liposomes:
Another form that can be used as
DNA vectors to transport normal
genes.
Liposomes are made out of lipids
and because of this, when they reach
the target tissue they fuse with the
cell membrane to deliver the DNA.
This has fewer side effects than
using viruses however but it is not as
effective.
Problems:
For gene therapy to be effective, a gene must reach the right place in the body
and become part of the normal workings of the cells involved.
It is essential to complete all steps correctly otherwise there may be dire
consequences. This includes, find the right gene, target the right cells in the
body, deliver the correct DNA of the required gene into these cells, make sure
the DNA is used correctly by these cells and that the procedure is performed
safely causing no injury or harmful side effects. Mistakes can cause damage that
can usually not be reversed.
For the gene delivery technology to the target cells to be effective, the new gene
must be inserted into enough cells to produce a change in the phenotype and
this can be difficult to achieve.
Gene therapy has been successful in treatment if only a few cells must be
altered such as stem cells in the bone marrow and in the retina. However gene
therapy is not as successful in treating large organs or tissues due to the large

volume of cells in these areas into which the gene must be inserted such as the
liver or lining of the respiratory tract. This is the difficulty in treating cystic fibrosis.
The treatment may also only be temporary if the receiving cells are rapidly
dividing and only have a short cell life.
The patient may have an immune response to the virus, therefore destroying the
vector (adenovirus)
The insertion of a new gene may trigger cancer-causing genes as genes
delivered by a retrovirus insert randomly into chromosomes and can cause
mutations.
Example:
Lebers Congenital Amaurosis is a genetic disease. Patients are missing a gene
in their retina cells which codes for an enzyme that allows them to turn light
energy into electrical signals
This causes degeneration of the retina and blindness.
In 2008 gene therapy successfully delivered the missing gene via a retrovirus
directly to the retina by injection.
All patients treated have seen such a large improvement in their sight and can
even read some lines on an eye chart.
Unfortunately only one eye was treated as scientists must establish whether
there are any long term side effects of the gene therapy
Process:
1) A cannula is pushed through the eye to the base of
the retina cells

2) Fluid containing a retrovirus with the normal gene is

injected beneath the retina

3) The fluid is absorbed by the pigment layer below

the retina. The virus infects the retina cells and
the enzyme needed for sight is expressed.

2. Process and analyse information from secondary sources to identify a current

use of gene therapy to manage a genetic disease, a named form of cancer or
AIDS

Severe Combined Immunodeficiency Disease X linked (SCID-X1):

SCID-X1 is a rare genetic disease with the defective gene found on the X
chromosome. As a result, this disease is more common in male infants.
The babies present to the doctor with chronic diarrhoea, persistent thrush,
pneumonia and sepsis.
This is because the stem cells in their bone marrow lack a receptor which
enables them to differentiate into Helper T cells so these babies have a greatly
reduced immune response.
They rarely survive longer than 1 year without a bone marrow transplant
To avoid infection, babies are kept in isolation boy-in-the-bubble.
This
Process:was the first disease to be successfully treated with gene therapy by French
scientists.
The normal human gene is
cloned in plasmids in
bacteria

Some stem cells are

removed from the babies
bone marrow

The DNA in a retrovirus

is modified. Only end
parts of the DNA remain

Human DNA is incorporated

into the virus (vector)

The virus is
introduced
into the stem
cell

Stem cells
multiply are
placed back
into the babies
bone marrow

Stem cells multiply

and the new DNA
is incorporated into
the chromosomes.

Unfortunately suitable donors cannot always be found and some babies may
reject the bone marrow.
18 children have now been treated with gene therapy for this disease. 15 are
healthy and are suggestive of a cure.
Unfortunately three have developed a leukaemia-like illness in 2003 as the gene
was randomly inserted next to a regulatory gene controlling leukaemia in these
patients. Research is ongoing to find a way to solve this problem.
Altering Immune Cells for Treatment of Advanced Melanoma:

Skin cancer is the most common of all cancers. A team of researchers at the
National Cancer institute (NCI) have demonstrated sustained regression of
advanced melanoma in a study of 17 patients by genetically engineering a
patients own white blood cells to recognise and attack cancer cells.

In a process called adoptive cell transfer, lymphocytes are first removed from the
patient with advanced melanoma. Next, the most aggressive tumour killing cells
are isolated, multiplied in the lab and then reintroduced to patients who have been
depleted of all remaining lymphocytes.

While reasonably successful, this method could only be used for melanoma
patients and only for those who already have a population of specialized
lymphocytes that recognize tumors as abnormal cells.

Thus, NCI researchers sought an effective way to convert normal lymphocytes in

the lab into cancer-fighting cells.
By infecting normal lymphocytes from a sample of blood from the patient with a
retrovirus in the laboratory, genes that encoded specific proteins, called T cell
receptors (TCRs), were into cells. When these genes are turned on, TCRs are
made and these receptor proteins decorate the outer surface of the lymphocytes
so that they activate the lymphocytes to destroy the cancer cells by binding to
certain molecules found on the surface of tumor cells.

In this study, newly engineered lymphocytes were infused into 17 patients with
advanced metastatic melanoma.
- Three patients who showed no delay in the progression of their disease had
improved the treatment of lymphocytes in the lab so that the cells could be
administered in their most active growth phase.
- Two patients experienced cancer regression, had sustained high levels of
genetically altered lymphocytes, and remained disease-free over one year. One
month after receiving gene therapy.
- All other patients still had 9 percent to 56 percent of their TCR-expressing
lymphocytes. There were no toxic side effects attributed to the genetically
modified cells in any patient.
Approaches to increase the function of the engineered TCRs include the
development of TCRs that can bind to tumor cells more tightly and further
optimised delivery methods using retroviruses are under investigation.
Researchers also believe it may be beneficial to further modify lymphocytes by
inserting molecules that assist in directing lymphocytes to cancerous tissues.
There is hope that this technology may be transferable to treat other cancers such
as breast and lung cancers.

Mechanisms of genetic change

1. Distinguish between mutations of chromosomes, including; rearrangements,
changes in chromosome number, including trisomy, and polyploidy and
mutations of genes, including, base substitution, frameshift

Mutation: Structural change to a chromosome that is a permanent, inheritable

change and where the chromosome state cannot be re-established or repaired.

Rearrangements:
---------------------See BLUEPRINT OF LIFE notes, Focus 4, Point 4-----------------------MUTATIONS OF GENES
Base Substitution:

This is where a single nucleotide and its partner or pair is replaced with a different
nucleotide pair. Base substitutions can result in:
1) No change in the amino acid and no effect on the protein (silent mutations)
- This occurs because more than 95% of DNA is non-coding so these mutations
have no genetic consequences.
- There is also more than one codon that codes for the same protein so no
apparent effect.
2) A change in the amino acid but no effect on the
protein
3) A change in the amino acid and an effect on the
protein
Missense mutations: base substations that do result in
a change to the amino acid.
E.g. Sickle cell anaemia
- This occurs when there is a base substitution in
the DNA that codes for the -globin protein.
- In normal -globin, the amino acid, glutamic acid is
produced from the codon GAG.
- However if a base substitution occurs that
exchanges the adenine with a thymine, the amino
acid valine is produced resulting in DNA coding for
sickle cell -globin.
Nonsense mutations: base substitutions that can change the codon for an amino
acid into a stop codon (UGA, UAA, UGA). This stops translation of the protein which
is inevitably shortened and almost always non-functional.
Frameshift Mutations:
These are mutations that shift the reading frame and is a result from a base
insertion or deletion.
E.g. THEBIGCATATETHERAT
THE BIG CAT ATE THE RAT
If a single letter is substituted (F for R) then sentence is slightly changed
THE BIG CAT ATE THE FAT
If a single letter is deleted (e.g. the G in BIG) this changes the reading fram so
that the sentence becomes comparatively meaningless
THE BIC ATA TET HER AT
An insertion or deletion that is one codon in length (three nucleotides) does not
result in a frameshift but will add or delete one amino acid.
E.g. Cystic fibrosis
CHANGES IN CHROMOSOMAL NUMBER:
Trisomy:
This can occur in meiosis, resulting in gametes with extra chromosomes or gametes
with missing chromosomes. This is called non-disjunction.

Non-disjunction can occur during the first or second stages of division in meiosis
The right side of the diagram
shows non-disjunction during the first
division, resulting in two gametes with 24
chromosomes and two gametes with 22
chromosomes.
The left side shows nondisjunction in the second division,
resulting in one gamete with 24
chromosomes and one with 22
chromosomes.
DIAGRAM:
Similar
to the cells produced
(note:
onlyabnormal
one pair of
in mitosis with
chromosomes
has been shown.)
numbers
of chromosomes,
many of the cells produced
in meiosis with too few or too many chromosomes will die.
If fertilisation does occur, many of the genes required for normal development
have been lost and the embryo will die.
However, a few embryos with alterations of their autosomal and sex
chromosomes do survive.
E.g. Down syndrome: (trisomy 21):
If the non-disjunction occurs in meiosis 1 then there is a 25% chance the parent
will have a down syndrome child. If the non-disjunction occurs at meiosis 2, then
there is a 50% chance of a parent having a child with down syndrome.
The enzyme expressed by this gene normally protects the cell from any harmful
substances being produced during normal cell metabolism.
However, in the case of children with three copies of the gene (trisomy 21), it is
thought that the extra production of this enzyme may cause an imbalance by
producing more than the normal quantity
of oxygen in the cells.
The excessive amount of oxygen
damages nerve and brain cells and the
cell membranes by destroying the lipids
in them.
This results in many of the abnormalities
seen in individuals with Down syndrome.
-------------------------------------------------See
Point 5----------------------------------------------Trisomy of sex chromosomes:
When non-disjunction occurs to the sex
chromosomes this can cause abnormalities.
Turner syndrome XO; female

One of the X chromosomes is missing or abnormal

Suffers from short stature and infertility, congenital heart defects, hearing
problems and absence of menstruation
Klinefelter syndrome XXY, XXYY, XXXY; male
Males born with two or more copies of the X chromosome.
Suffers from learning difficulties, a tendency to grow at a slightly quicker rate than
others so final height is more than expected (but in normal range), reduction in
body hair, bead growth and testicular size, can be infertile.

Polyploidy:
Most cells in the human body are diploid (2N), with 46 chromosomes, but some
cells such as those in the ovaries and testes are haploid (1N), with 23
chromosomes.
Some individuals can be monoploid meaning they have only one instead of two
sets of chromosomes.
These organisms normally develop from unfertilised eggs
E.g. ants, bees, water fleas.
Polyploidy is when an organism has cells with three or more sets of
chromosomes (3N or more).
It is rare in animals but occurs more commonly in plants.
E.g. wheat (hexaploid (6N)), strawberries (octaploid (8N))
Polyploidy occurs because of non-disjunction at the first or second meiotic
division because the spindle apparatus is not functioning. If the resulting gamete
has two sets of chromosomes and is fertilised by a gamete with a normal set of
chromosomes, then a triploid organism will result.
Polyploidy often confers advantages in plants such as higher yield (so better
economically), greater chance of survival (has more seeds to pass on), less
space (so better environmentally).
This is rarer in animals than in plants because most polyploid animals fail to
develop. Some types of polyploid fish and insects have been discovered.
However polyploidy is often lethal in humans.
2. Outline the ability of DNA to repair itself
Damage to DNA is fairly common, but the genome has the ability to repair itself.
A mutation only occurs when the repair system fails to work properly
There are more than 130 genes which are responsible for repairing DNA and
these genes operate during DNA replication to repair spontaneous damage to the
DNA caused by chemical reactions in the nucleus, chemical mutagens and
radiation.

 The types of molecules involved in DNA repair depend on the type of damage that
has occurred.

Damage Reversal:
During replication the enzyme DNA polymerase
proof reads the base order.
It repairs any copying errors by using the
undamaged strand of DNA in the double helix as a
template to fix and replace the
incorrect damage base sequence

Damage removal:
Damage affecting longer strands of
DNA uses an excision repair system
that typically involves three enzymes:
One to remove the damaged
section
One (DNA polymerase) to add the
missing bases
One to repair the break in the
backbone (DNA ligase)

Failure to repair:
The rate of DNA repair is dependent on cell type, age of the cell and the extracellular
environment. A cell that has accumulated a large amount of DNA damage or can no
longer effectively repair the damage can
Enter a stage of irreversible dormancy (senescence)
commit suicide (programmed cell death or apoptosis)
Commence unregulated cell division which can lead to cancer.
3. Describe the way in which transposable genetic elements operate and discuss
their impact on the genome
In 1944 while studying the genetics of corn, Barbara Mc Clintock observed that
coloured spots on kernels would sometimes appear in one generation and
disappear in the next.
She discovered that the differences corresponded with damage to one pair of
chromosomes and hypothesised that genes were being transposed (moved) from
one part of the chromosome to another.

This was also evident when there was a discovery that genes in bacteria
appeared to change position.
This is called transposable genetic elements
Transposons are segments of DNA which code for
an enzyme that can move the segments directly
from one part of the chromosome to another. They
move by a cut and paste or copy and paste
mechanism.
A transposon may stay in one position for several
generations and then suddenly move to a new
location. Sometimes it may be inserted into another
gene where it can cause mutation.
Transposable elements make up 20 25% of the
mammalian genome. Transposons can disable genes,
cause uneven crossing over and leave incorrectly
repaired
E.g.
Indian genes.
maize (corn)
The been
purpleimplicated
kernels occur
where the
cells
have the
Theyhave
in diseases
such
as haemophilia,
normal
C allele.
Duchenne
muscular dystrophy and porphyria.
White kernels occur where the C allele is disrupted by the presence of a
transposon.
In the variegated kernels, the white areas have the disrupted gene, while in the
purple areas the transposon has moved and gene has reverted to the normal C
allele.
4. Distinguish between germ line and somatic mutations in terms of their effect
on species
Somatic Cells:
Mutations that occur in somatic cells (body cells) are not passed on to offspring, it
only affects the individual.
If a somatic mutation occurs in a cell during the first few cell divisions after
fertilisation it can lead to a mosaic an organism with some mutated cells and
some normal cells.
If a mutation occurs in a somatic cell after birth, it may produce cancer (e.g. skin
cancer forming as a result of prolonged exposure to UV radiation)
Germ Cells:
Mutations that occur in germ cells (that form gametes) can be passed onto
offspring and cause other mutations.
They affect the sperm or ova and the offspring produced are known are mutants
The effect this has on a species is to produce sources of variation, and if the
mutant has an advantage to the species it can become the most common
characteristic or trait, through the process of natural selection.
E.g. Down syndrome (------------------see point 5------------------)
5. Process and analyse information from secondary sources to describe the
effect of one named and described genetic mutation on human health

Name of Genetic
Mutation on
Down Syndrome (trisomy 21)
Human Health
- During meiosis, the pairs of chromosomes line up in the centre of the cell and
Cause
attach to spindle fibres. The event of dividing and travelling to the poles of the
dividing cell so they are contained as the cell divides is called disjunction.
- Occasionally one pair of chromosomes does not divide and so the whole pair
travels to one end of the dividing cell. This means that in the resulting cells, one
will have 24 chromosomes and the other will have 22 chromosomes. This
accident is called non-disjunction. (---See Point 1---)
- If a sperm or egg with an abnormal number of chromosomes merges with a
normal mate, the resulting fertilized egg will have an abnormal number of
chromosomes. In Down syndrome, one cell has two 21st chromosomes instead
of one, so the resulting fertilized egg has three 21st chromosomes.
- - Recent research has shown that approximately 90% of the abnormal cells are the
eggs. Research is currently aimed at trying to determine the cause and timing of
non-disjunction
maternal age.
the
Poor
muscle tone error, but there is definitely connection with
Effect on
Individual
Slanting eyes with folds of skin at the inner corners (called epicanthal folds)
Short, broad hands with a single crease across the palm on one or both hands
Flat bridge of the nose
Short, low-set ears
Short neck
Small head
Individuals with Down syndrome are usually smaller than their non-disabled
peers, and their physical as well as intellectual development is slower.
Process of
Diagnosis

Down syndrome of an individual is often diagnosed shortly after birth.

The diagnosis of Down's syndrome is usually made soon after birth because of
some subtle differences in the baby's appearance. Then the baby has a
chromosome test called a karyotype. It usually takes a few days to get the results
of this blood test
Down Syndrome is increasingly being diagnosed during pregnancy (prenatally) due
to abnormal ultrasound findings, an abnormal result on a blood test done or by
chorionic villi sampling (CVS) which is a test done on the placenta between 10 and
12 weeks of pregnancy.

Treatment

There is no cure to this disease. However individuals with down syndrome are
more prone to:
- Eye problems
- Congenital heart disease
- Seizure disorders
- Leukemia and other cancers
- Developmental delay
- Immune system problems
- Mental retardation
- Thyroid problems
- Bone, muscle, nerve, or joint problems - Premature aging
- Alzheimer's disease
- Hearing problems
Therefore treatment includes:
- Surgery
- Regular checkups and screening
- Counseling and support
- Medications

Prognosis

Earlier detection
could assist in
the treatment of
the disease

Because of these medical conditions and their associated complications, a person

with Down syndrome is at increased risk of premature death and there is an
increased risk of vision and hearing problems, infections, and cancer in people with
Down syndrome.
Allows the choice of parents to abort the pregnancy

Selective breeding is different to gene cloning but both processes may change
the genetic nature of species
1. Explain, using an appropriate example from agriculture, why selective
breeding has been practiced
-----------------------------See Blueprint of Life: Focus 2, Point 10------------------------------Selective breeding (or artificial selection) is practiced as a way of improving the
quality and yield of farm animals and food crops. This can be thought of as the
manipulation of the phenotype in the offspring to achieve desirable characteristics.
This result of this is that these species can:
Produce crops that give high yields and increase nutritional value
Resist disease
Show tolerance to drought and cold
Improve productivity by developing faster growing and larger animals
Produced good quality food in larger quantities.
2. Describe what is meant by gene cloning and give examples of the uses of
gene cloning
Gene cloning: a process which isolates a single gene so that multiple identical
copies can be made. This is done by inserting the gene into a bacteria plasmid
and then allowing the bacteria to replicate the gene (recombinant DNA
technology).
DNA gene splicing:

-------------------------See Blueprint of Life Notes: Focus 5, Point 2---------------------------3. Distinguish between gene cloning and whole organism cloning in terms of the
processes and products
Gene Cloning

Whole Organism Cloning

Aim

To produce genetically identical

copies of a gene

To produce genetically identical copies

of an organism

Products

A single gene, and sometimes the

products of that gene (the protein)

The whole genome. A whole organism

which contains trillions of cells

Numbers usually
cloned at a time

Huge numbers billions or trillions

Small numbers, often just one

Methodology
(processes)

The gene to be cloned is cut out

and added to the DNA of another
organism and then replicated

A variety of techniques including tissue

culture in plants and somatic cell
nuclear transfer in animals.

4. Discuss a use of cloning in animals or plants that has possible benefits to

humans
Cloning of animals for agricultural purposes is a genetic refinement of selective
breeding.
By cloning, the results are identical to the donor cell, whereas in selective breeding
the resultant offspring may contain some unwanted genes as well as the desirable
ones.
The time involved in producing these desirable cloned populations is far less than
that involved in selective breeding.
Advantages:
Similar to those of selective breeding where large numbers of plants or animals
could be bred for factors such as:
o more meat
o finer wool
o more milk
o higher crop yield
\

o resistance to disease.
Disadvantages:
All members of a population being genetically identical would mean that a lack of
resistance to a new strain of disease or a change in environmental conditions could
decimate the whole population as there is no diversity within the population.
5. Analyse and present information from secondary sources to trace the history
of the selective breeding of one species for agricultural purposes and use
available evidence to describe the series of changes that have occurred in the
species as a result of this selective breeding
Belgian Blue
The Belgian Blue has been selectively bred to obtain the
double muscling gene. This gene allows the muscles to
grow to a larger size then the average cow. The natural
mutation of the gene codes for myostatin, a protein that
counteracts
muscle
growth.
As the
enzyme or
is unable to
6. Identify data
sources,
choose
equipment
function
at its gather,
capacityprocess
the result
is lean
muscle
growth.
resources,
and
analyse
information
from secondary sources to describe the processes
used in the cloning of an animal and analyse the
methodology to identify ways in which scientists
could verify that the animal produced was a clone
Dolly was a clone of a Finn-Dorset ewe. A nucleus containing an entire set of
chromosomes from the udder set of the Finn-Dorset ewe was removed. An
unfertilised egg from a Scottish Blackface ewe was also removed and
evacuated.
The nucleus was fused into the egg, and placed into a culture containing the
early embryo. This was then planted in a surrogate Scottish Blackface ewe
mother.
The embryo developed and exhibited the same characteristics as the FinnDorset ewe.
Methods that were used by scientists to verify Dolly was a clone included:
- DNA fingerprinting, to compare DNA fragments with the original and clone sheep
- DNA Hybridisation
- Intron sequences were compared with the original and clone sheep, as each
organism has unique intron sequences.
The timing of gene expression is important in the developmental process
1. Identify the role of genes in embryonic development
On fertilisation, two haploid sets of chromosomes join to produce a zygote
containing all the genes necessary to grow into a new individual.
In the pre-embryonic period the zygote divides by mitosis to become a hollow ball
called a blastula.

This ball then embeds in the wall of the uterus in implantation and following this, is
gastrulation which is the formation of the three germ layers of the blastula the
ectoderm, endoderm and mesoderm. From this all tissues and organs form.
Gastrulation is important as if this did not occur, specialised organs could not be
formed.
Patterning is a process where genes cause embryonic cells to differentiate into
body regions and allow the cells to become specialised in their size, shape and
function. In the cells specific genes become switched on so the cell produces
particular proteins to allow this.
The activities of genes are regulated by other genes to control all cell activities,
production of proteins and enzymes, at different times of development.
A variety of proteins, activators and repressors, link up with different parts of DNA
to cause interaction between various other proteins and genes. The result is that
some genes are affected and expressed and others are repressed or inhibited.
Genes are turned on or off according to their position in the embryo, function and
age of the developing embryo/foetus/individual.
2. Summarise the role of gene cascades determining limb formation in birds and
mammals

A gene cascade is a series of genes turning on and off in the correct sequence of
gene expression during the development of an embryo

Morphogens are chemicals that control which genes are switched on/off and thus
control the future identity of a cell. This is important in limb development as it
determines the cells specialisations in the limb

AER is the apical ectoderm ridge. This is the region near the tip of the bud that
controls limb length

AER control of limb development is shown when it is removed, no limb

development or when another is added, two limbs will grow.

3. Describe the evidence which indicates the presence of ancestral vertebrate

gene homologues in lower animal classes
Homologous structures such as pentadactyl limbs in vertebrates provide evidence
for evolution from common ancestors.
DNA sequences that are similar in many organisms are called homologue genes,
homeobox or Hox genes. These genes regulate the development of an organism
by producing proteins that switch other genes on and off.
E.g. The gene cascade for skeletal and neurological development in limbs is
similar in organisms such as humans, chickens, rodents, insects, nematodes and
molluscs. This has become evident through experiments that have confirmed that
the homologue gene from an amphibian can regulate the corresponding gene in
mammals. A mammal homologue gene can regulate the corresponding gene in
insects such as fruit flies.

These homologue genes exist in many eukaryotic organisms, both vertebrates

and invertebrates as well as some fungi and plants and suggests common
ancestry between all eukaryotic organisms
4. Discuss the evidence available from current research about the evolution of
genes and their actions
Homologues, homeotic or Hox genes are found in most or all groups of
multicellular animals and show similar DNA sequences suggesting that these
genes evolved in a common ancestor. These genes, being similar in both
structure and function, are expressed in similar sequences on chromosomes.
The study of DNA sequences provides evidence for the evolution of genes.
Sequences of bases in genes that do not change or change very slowly over time
are used to measure relationships between groups of organisms. For example, in
mice, the gene for the development of eyes is similar to the gene in insects.
The study of mutations of homeotic genes shows that a small mutation can
suddenly affect an organism as a gene cascade is altered. A mutation in a
homeotic gene can cause one part of the body to develop into another. For
example, in Drosophila fruit flies, one mutation in a homologue gene can result in
legs growing on the head rather than antennae. In humans, a mutation in a
homeotic gene for the development of the bones of the skull results in rigidity in
the joints between these bones. This can result in abnormal brain growth and
possible mental retardation. The base sequence for this gene is very similar in
many animals.
These gene changes can then result in often dramatic alterations in organisms
which may then lead to the rapid evolution of new body structures. This is
evidence for the expansion of diversity of living things and the theory of
punctuated evolution.
5. Identify data sources, gather, process and analyse information from secondary
sources and use available evidence to assess the evidence that analysis of
genes provides for evolutionary relationships
--------------------------------------------See Points 3 and 4-------------------------------------------