Further inheritance

1. Chi squared (X2) test is a statistical test that can be used to compare
your observed results with expected(theoretical) results and determine if the
difference between observed and expected results is due to chance or not.

The Chi2 test is used when:

• the data is nominal – i.e. can be given categories such as different phenotypes
• you have actual observations rather than percentages or fractions
• you can calculate expected values, e.g. based on a Mendelian ratio.

Before calculating a Chi2 statistic you need to formulate a null hypothesis. This states
that:

there is no statistically significant difference between the observed and the

expected results.

You then follow these steps:

1. Decide how many sets of data (number of different phenotypes) you have.

=n

2. Calculate how much the sets can vary.

This is called the degrees of freedom.

(df) = n-1

3. For each set of data calculate:

the expected results = E

the observed – expected = O-E

the (observed – expected)2 = (O-E)2

divide the (observed – expected)2 = (O-E)2

by the expected E
add these numbers together to give = Σ(O-E)2
the Chi2 statistic E

Once you have calculated a value of Chi2 you need to compare your calculated value
against the table of Chi2values. The table of Chi2 values helps you to decide on
the probability that your observed results are down to chance alone, or if there is a
significant difference between them and the expected results.

You find the critical value for Chi2 by using a probability table; you find the number of
degrees of freedom and the 0.05 probability and this gives you the critical value for
Chi2.

Low Chi2 = theoretical result close to real result

Higher Chi2 = theoretical result not close to real result => different is not due to anything by chance => there
has to be something else to account for the difference (e.g. linkage, lethal allele combination)

If your value of Chi2 is less than the critical value at p = 0.05 you accept the null
hypothesis – there is no significant difference between observed and expected
values, and the differences between observed and expected values are due to chance
alone, e.g. random fertilisation.

If your value of Chi2 is equal to or greater than the critical value at p = 0.05,
you reject the null hypothesis – there is a significant difference between observed
and expected values, and the differences between observed and expected values
are not due to chance, e.g. linkage.

Example:
 2. Muta5ons
A mutaBon is a random, spontaneous change in the amount, arrangement or
structure of the DNA of an organism.
The rate of mutation can be increased by ionising radiation and mutagenic chemicals
(Mutagens) such as:

• X-rays, gamma radiation and UV light

• chemicals, such as polycyclic hydrocarbons in cigarette smoke.

• A mutagen which causes cancer is a carcinogen.

 a. Gene MutaXon
Gene mutaXons are changes in the base pairs within the genes. They can take the
form of duplicaBon, inserBon, deleBon, inversion or subsBtuBon of bases. They occur
during DNA replicaXon before cell division.

Example:
CCT GAG GAG may change to CCT GTG GAG
CCT GAG GAG may change to CCA TGA GGA G
CCT GAG GAG may change to CCG AGG AG.

Base addi5on or dele5on usually have a significant effect on polypep5de that the allele
codes for.

• A mutation in an intron (non-coding region of a gene) has no effect as introns

are spliced out during the formation of functional mRNA.
• A silent mutation a mutation that has no apparent effect on an organism.
• A missense mutation changes the triplet code and the amino acid incorporated
into the protein. This can change the way that a polypeptide chain folds and
can affect its functionality.
• A nonsense mutation introduces or deletes a stop codon, thus changing the
length of the polypeptide chain.

Reasons for silent muta5on:

many amino acids have more than one triplet code
muta5on in an intron
some5mes even if there is a change in one amino acid(missense mutation), the structure
can be largely unaltered.

Case Study: SICKLE-CELL ANAEMIA

A gene mutation (substitution) in the gene producing haemoglobin results in a defect
called sickle-cell anaemia.

The DNA base triplet codes for glutamic acid are CTT or CTC. Two of the codes for
valine are CAT and CAC. In either case, the substitution of A for T as the second base
would bring about the formation of haemoglobin S.

The mutant gene is co-dominant in terms of protein expression. In the homozygous

state for haemoglobin S, the individual suffers the disease and rarely survives longer
than a few years after birth.

In the heterozygous state, most people show no symptoms. (So some people say the
disease is recessive) However, 30-40% of red blood cells are sickle cells - the rest are
normal. This is called sickle-cell trait. A heterozygous person has reduced ability to
absorb and transport oxygen, but this reduces the risk of being infected with malaria.
Sickled red blood cells are more likely to get stuck in capillaries and can lead to low
oxygen supply to tissues, and clot formation.

b. Chromosome mutaXon

CHROMOSOME MUTATIONS can occur when chromosomes are damaged and

break, or there is a change in the number of chromosomes.

If chromosomes break, they will normally repair themselves (the DNA will re-join), but
they may not repair themselves correctly. This can lead to big changes in the
structure of the DNA and may affect a large number of genes.

Changes in chromosome numbers

This is usually the result of errors occurring during meiosis. This happens at anaphase
I or II when homologous chromosomes (anaphase I) or chromatids (anaphase II) fail to
separate correctly as shown in the diagram.

Aneuploidy is the loss or gain of a single chromosome. It results in the formation of

gametes with either one too many or one too few chromosomes.

In the example shown here, during meiosis to form gametes, one of the pair of
chromosomes 21 did not separate at anaphase and so the pair moved to one pole of
the cell - this is called non-disjunction. The gamete formed had two copies of
chromosome 21.

After fertilisation, a zygote formed from one gamete with a single copy of
chromosome 21 and another gamete with two copies of chromosome 21. This means
that the embryo has an extra chromosome and is said to be trisomic. Down
syndrome is the result of having three copies of chromosome number 21.

In Down syndrome, the error occurs in the production of ova rather than sperm and
the incidence of the mutation is related to the age of the mother.

Aside:

Polyploidy involves changes in the number of whole sets of chromosomes and

has played an important role in the evolution of many plants.

For an organism to be fully fertile, its chromosomes must be in homologous

pairs and be able to form bivalents during prophase I. If there are uneven/odd
numbers of chromosomes, bivalents cannot form, meiosis does not continue,
and no gametes are produced.

Example: mule
Another form of chromosome mutation is Changes in chromosome structure

During chiasmata formation, mistakes can occur when chromatids break at these
points and re-join with the corresponding pieces of chromatid on its homologous
partner.

Sometimes, non-homologous chromosomes also pair up during prophase I and

crossing over results in part of one chromosome being transferred to another.

One example of this is the transfer of the SRY gene from a Y chromosome to an X
chromosome due to an error in crossing over during prophase I. The SRY gene
controls whether the developing embryo secretes testosterone and its genitalia
develop to become male testes.

An individual with two X chromosomes – therefore female – will then produce

testosterone as an adult and may have some male characteristics. Conversely, a male
with no SRY gene develops into an XY female.

c. Somatic mutation vs germline mutation

Somatic mutation: may cause cancer
Germline mutation: affect offspring

Mutations can be useful in a population level: increasing genetic variation

Beneficial mutations may appear: giving selective advantage

d. Oncogene: An oncogene is a gene that, when mutated or expressed at high

levels, can turn a normal cell into a tumour cell.

Many cells normally undergo a programmed form of death called apoptosis. Activated
oncogenes can cause those cells to survive and increase in number instead.

Most oncogenes require an additional step, such as a mutation in another gene, or

environmental factors, such as a viral infection, to cause cancer (e.g. cervical cancer
being triggered by HPV infection).

Proto-oncogenes are genes that if they mutate can become oncogenes.

Tumour suppressor genes are different to oncogenes and proto-oncogenes in that

they are protective genes that control cell growth by suppressing cell
division. Oncogenes increase the rate of cell division. Mutations to both types of
genes can cause cancer.
 3. Epigene5cs
EpigeneBc changes do not affect the sequence of bases – therefore they
are not mutaBons. However, the changes may be heritable and can be passed
on to your offspring or they can affect you alone.

Genes are expressed when they are copied into mRNA

during transcripBon and then translated into polypepXdes. Anything that
affects how a gene is transcribed can affect the phenotype.
For example:

• if a tumour suppressor gene is silenced there is no control over cell growth

and division which can lead to cancer development
• overexpression of an oncogene can lead to increased rates of cell division and
again cause cancer.

a. DNA Methylation

Methyl groups (–CH3) are found in many foods, medicines and other
drugs, and in some pollutants. They can become free radicals which are
highly reactive. Methyl groups can be added to cytosine bases.

Every gene has a sequence of bases before the first exon, called its promoter region.

Generally, if more methyl groups are added to the promoter region of a gene, it is less
likely that the gene will be expressed.

Methylation of DNA can stop proteins binding to the DNA that are needed to start
transcription. This means that the more cytosine bases that are methylated, the
fewer transcription proteins can bind to the gene and less mRNA is produced.
 b. Histone acetyla/on
Chromosomes are a combination of DNA wrapped around protein complexes
called histones. The combination of DNA and histones is called chromatin. The
organisation of chromosomes is shown in the diagram.

The histones can be modified by the addition or removal of acetyl/ethyl groups

(CH3CH2--).

• If more acetyl groups bind to the histones, the DNA unwinds and becomes less
compact. This is called euchromatin.

The genes in euchromatin or open chromatin can be transcribed – i.e. they can
be expressed.

• If fewer acetyl groups bind to the histones the DNA remains compact and
tightly wound around the histones. This is called heterochromatin.

The genes in heterochromatin cannot be transcribed – they are not expressed.