Chapter 3 Bioplanet T.

Rami Mawasi

Mutation

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Chapter 3 Bioplanet T.Rami Mawasi
Mutation and the environment

Mutation is a change in the nucleotide sequence. A mutation is a spontaneous event capable of producing
new phenotype from an already existing one.
Transmission of mutation:
Mutations in the germ cells can be transmitted; in
contrast mutations in the somatic cells are not
transmitted to the offspring
Example for somatic cell mutation:
An over exposure to UV-B rays may cause mutations in
the cells of exposed skin areas.
1. Pick out the role of the DNA repair proteins.

2. Pick out the two causes that lead to the

formation of cancerous cell.

3. Specify if this mutation transmitted to offspring.

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Types of mutation

There are three types of gene mutation (substitution, insertion and deletion). A change in the DNA
sequence alters the corresponding sequence of mRNA and may alter the corresponding protein.
(Note: the change of only one nucleotide is called point mutation)

There are types of mutation

1. Substitution.
2. Deletion
3. Insertion.

Note that:
The function of the protein depends on:
A. 3D Structure
B. Sequence

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E ect of
Type of mutation Normal Mutant
mutation

…CCA-GAG-ACT… …CCA-GTG-ACT… Missense,

DNA
5 6 7 5 6 7 altered
polypeptide.
Substitution mRNA …CCA-GAG-ACU… …CCA-GUG-ACU… Change in
sequence and
Protein ….pro - Glu - Thr …. pro - Val - Thr 3D structure

…CCA-GAG-ACT… …CCA-GAA-ACT…
DNA
5 6 7 5 6 7
Silent, functional
Substitution
mRNA …CCA-GAG-ACU… …CCA-GAA-ACU… protein

Protein ….pro - Glu - Thr …. pro - Glu - Thr

…CCA-GAG-ACT… …CCA-TAG-ACT… Nonsense,

DNA
5 6 7 5 6 7 incomplete
polypeptide.
Substitution mRNA …CCA-GAG-ACU… …CCA-UAG-ACU… Change in
sequence and
Protein ….pro - Glu - Thr …. pro - stop 3D structure

Deletion

E ect of
Type of mutation Normal Mutant
mutation

..TAC-ACC-ACG-A.. ..TAC-CCA-CGA.. Frameshift,

DNA
5 6 7 5 6 7 altered
polypeptide.
Deletion mRNA UAC-ACC-ACG-A.. ..UAC-CCA-CGA.. Change in
sequence and
Protein …Tyr - Thr - Thr… …. Tyr - Pro - Arg 3D structure.

Insertion

E ect of
Type of mutation Normal Mutant
mutation

..TAC-ACC-ACG-A.. ..TAC-GAC-CAC-GA.. Frameshift,

DNA
5 6 7 5 6 7 altered
polypeptide.
Insertion mRNA UAC-ACC-ACG-A.. ..UAC-GAC-CAC-GA.. Change in
sequence and
Protein …Tyr - Thr - Thr… …Tyr - Asp - His 3D structure.

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Based on the previous cases:
1. “Mutation is not always harmful”. Justify this statement.
2. Compare the sequence of normal and mutant alleles.
3. Indicate the position of mutation in each case.

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Genes and Multiple alleles:

Genetic polymorphism: means the presence of many

functional alleles of the same gene.
An example for the multiple alleles is human ABO
blood group system.
This system is defined by the presence of O, A and B
molecules on the surface of red blood cells. These
molecules have the basic structure called substance
H, but they differ by the absence or presence of
supplementary sugar.
The gene coded for substance H found on
chromosome 19 and has two alleles H and h, where
H>h.
If an individual has hh genotype this indicates he has
no substance H at the surface of RBC's.
- Allele A is coded for enzyme transferase A
that catalyzes the addition of N-
acetylgalactosamine to substance H

- Allele B is coded for enzyme transferase B

that catalyzes the addition of galactose to
substance H

- Allele O is coded for enzyme transferase O

which is a truncated and inactive enzyme and
does not add any sugar to substance H.

The gene coded for ABO blood groups found on

chromosome 9.

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Major histocompatibility complex (MHC)
In human it is also designated by human leucocyte antigen (HLA). This HLA is coded by 6 genes
that are located on chromosome 6. These 6 genes are DP, DQ, DR, B, C and A. Loci B, C and A
are coded for HLA class I which is located on the surface of all nucleated cells, however loci DQ,
.DP and DR are coded for HLA class II that are found on the surface of some immune cell
As shown in the adjacent document that the
genes coded for HLA are highly polymorphic
and it is rare, except for identical twins, to
.have to individuals with the same HLA
Note that the 6 genes are absolutely linked
together and inherited as a haplotype from the
.parents to the descendants

Note: RBC’s have no HLA on their membranes since they are enucleated cells; instead they have
.antigens
:β globin gene

β-globin gene is an example of polymorphic gene among the population. It is coded for β-globin
.polypeptide

The document above shows the non-transcribed strand for the gene coded for normal β-globin
.polypeptide

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The adjacent document shows examples
of diseases resulted from the mutation in
.the gene of β-globin
Note that the severity of the diseases
depends on whether one or both alleles
are mutant and on the location of this
.mutation in the gene

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Restriction enzymes

These enzymes are extracted from bacteria and they have the ability to cut the DNA. Each restriction
enzyme cut the DNA at a specific site (restriction site or cleavage site), this restriction site located in a
recognition site.
For example, the restriction enzyme Eco RI recognize
the sequence:

This sequence is considered as recognition site. When

Eco RI recognizes this sequence it cuts between G and
A.
The table below shows number of restriction enzymes
and their effect on the DNA:

In the adjacent document we add two different restriction

.enzymes on the same DNA molecule
?How many restriction fragments do we obtain in each case -

Indicate the number of restriction sites and the number of -

.recognition sites, on the DNA molecule, for each enzyme

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Gel Electrophoresis

Procedure:
1. Cut the DNA with restriction
enzyme.

2. Put the restriction fragment in the

well.

3. Apply electric current.

The fragments will migrate from

the negative pole to the positive
pole since the phosphate group in
the DNA imparts a negative
charge on the DNA molecule.
4. Add ethidium bromide which stain
the fragments and allow the
visualization of the bands since it is fluorescent under UV light.

5. Observe under UV light.

Notes:
- The distance a fragment migrates depends on its size, where the larger fragments migrates
slower.

- If two fragments
have the same size,
they will reach the
same point.

- The size of the

fragments is
measured in base
pair (bp) or kilo
base pair (kbp).

- The base sequences

of the DNA vary
from one allele to
another.

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- Consequently, if the mutation occurs within the recognition site, the restriction enzyme
will cut two different alleles of the same gene at different sites and thus yields fragments of
different sizes and numbers.

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Restriction fragment length polymorphism: (RFLP)
Mutation in the coded region of the genome usually affect the phenotype and can be detected by the
variation of the phenotype. However, mutations occur in the non-coding regions do not affect the
phenotype, and hence can only be revealed by the variation in the restriction maps of the region in
question. This is because the restriction map is independent of gene function.
A difference in the restriction map between two individuals is called restriction fragment length
polymorphism (RLFP).
So the genetic polymorphism can be detected by phenotypes and RLFP analysis.

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Localization of a sequence or gene on a chromosome

if you have to localize a gene or a sequence of nucleotides on a chromosome so you must know the exact
sequence of this gene.
We follow the steps of Fluorescence in situ
hybridization (FISH technique).
- First of all, we make a probe for the
gene to be located.

- Denaturation (separation of the two

DNA strands by heating)

- Add the probe

- The probe will bind to its complement

strand (hybridization)

- Washing

- Observe under fluorescent microscope.

Characteristics of the probe:

- The probe is a single stranded synthetic molecule.

- The probe must be complement to the sequence or gene to be located.

- The probe is either radioactive or fluorescent.

Genetic Map:
FISH technique allow scientists to construct the genetic map i.e. to
localize all the genes on the chromososmes.

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DNA fingerprint:

Jeffrey’s technique:
1. Cut the DNA with restriction enzyme.

2. DNA fragments are separated by gel electrophoresis.

3. The fragments are then transferred into solid membrane; this technique is called “southern
blotting”.

4. Denaturation for the fragments.

5. The radioactive probes are added.

The probe is complement to a DNA sequence that occurs frequently throughout the
genome (multi-locus probe)
6. Washing, to eliminate un-binded probes.

7. autoradiography to observe the bands.

the adjacent document shows the DNA fingerprint

for two identical twins and that of their sister.

Paternity test:

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Extraction from official exams

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Official exams

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2024/1st session

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2024/2nd session

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2022 second session

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2021 1st session

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2017 2nd session

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2013 2nd session

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2011 2nd session

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2005 2nd session

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Sample by
T.Rami Mawasi

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