GENETICS

SEX DETERMINATION

Sexual reproduction is the formation of offspring that are genetically distinct from their
parent; most often the parent contributes genes to their offspring. Among most
eukaryotes, sexual reproduction consists of two processes that lead to an alternation of
haploid and diploid cells: meiosis produce haploid gametes and fertilization produces
diploid zygotes. The term sex refers to sexual phenotype. Most organisms have only two
sexual phenotypes: male and female. The fundamental different between males and
females is gamete size: male produce small gametes: female produce relatively large
gametes. The mechanism by which sex is established is termed sex determination. Sex
determination is a process of sex differentiation which utilizes various genetical concepts
to decide whether a particular individual will develop into male or female. We defined
the sex of an individual in term of the individual’s phenotype – ultimately, the type of
gametes that is produces. Sometimes an individual has chromosome or genes that are
normally associated with one sex but a morphology corresponding to the opposite sex.
For instance, the cells of female human normally have X chromosomes, and the cells of a
males have one X chromosome and one Y chromosome. A few rare persons have male
anatomy, although their cells each contain two X chromosome. Even though these people
are genetically female, we refer to them as male because their sexual phenotype is male.

There are many ways in which sex differences arise. In some species, both sexes are
present in the same individual a condition termed hermphroditism, organisms that bear
both male and female reproductive structures are said to be mioecious (meaning “one
house”). Species in which an individual has either male or female reproductive structure
are said to be dioecious (meaning “two house”). human are dioecious. Among dioecious
species, the sex of an individual may be determined chromosomally, genetically, or
environmentally.

The various mechanisms of sex determination are:

i. Chromosomal sex determination

ii. Genic balance mechanism
iii. Male haploidy or Haplodiploidy mechanism
iv. Sex determination due to environmental factors

I. CHROMOSOMAL SEX - DETERMINING SYSTEMS

1
The chromosome theory of inheritance states that genes are located on chromosomes,
which serve as the vehicle for gene segregation in meiosis. Definitive proof of this theory
was provided by the discovery that the sex of certain insect is determined by the presence
or absence of particular chromosomes.

The chromosomes, which have no relation with sex and contain genes, which determine
the somatic characters of an individual are known as autosomes. These chromosomes do
not differ in morphology and number in male and female sex. Those chromosomes,
which differ in morphology and number in male and female sex and contain genes
responsible for the determination of sex are known as allosomes or sex chromosomes.

Differences between Autosomes and Allosomes

AUTOSOMES SEX CHROMOSOMES

Refer to other than sex chromosomes. These are sex chromosomes.

Morphology is similar in male and female Morphology is different in male and female sex.
sex.

The number is same in both the sexes. The number is sometimes different in male and
female sex.

Generally control traits other than Usually determine sex of a sex.

individual.

Number of autosomes differs from species Each diploid organism usually has two allosomes
to species.

Do not exhibit sex linkage. Exhibit sex linkage.

The chromosomal influence on sex, in certain insects, has been shown for the first time
by McClung in 1902 to be associated with a special sex determining ‘X’ chromosome.
McClung proposed that a male had one ‘X’ chromosome per cell (XO) and a female has
two ‘X’ chromosomes (XX). Later Stevens and Wilson (1905) found same number of
chromosomes in both sexes of milk weed bug. In females all chromosomes were paired
and the homologues were equal in size (homomorphic). In the male, all the chromosomes
were paired, but the chromosome identified as homologous to the “X” Chromosome was
distinctly smaller and was called the “Y” Chromosome (Heteromorphic).

2
Thus, allosomes are generally of X and Y types, but in birds they are of Z and W types.
Sex with similar type of sex chromosomes (XX or ZZ) is known as homogametic sex and
with dissimilar type of sex chromosomes (XY or ZW) as heterogametic sex. These are
two types: a) Heterogametic male and b) Heterogametic female.

a) Heterogametic male: In this mechanism, the female sex has two ‘X’ chromosomes,
while the male sex has only a single ‘X’ chromosome. As the male lacks a ‘X’
chromosome during meiosis, 50% of the gametes carry ‘X’ chromosome, while the rest
do not have the ‘X’ chromosome. Such a mechanism, which produces two different types
of gametes in terms of sex chromosome is called heterogametic sex. The female sex here
is called homogametic sex because it produces similar type of gametes. The
heterogametic male may be of the following two types.

i) XX – XO

ii) XX – XY

i) XX - XO: In certain insects belonging to orders Hemiptera (true bugs), Orthoptera

(grass hoppers) and Dictyoptera (cockroaches), female has two ‘X’ chromosomes (XX)
and are, thus homogametic, while male has only single ‘X’ chromosome (XO) There is
no O chromosome: the letter O signifies the absent of a sex chromosome. This
mechanism was found by C.E. McClung in 1902. The presence of an unpaired X
chromosome determines the masculine sex. The male being heterogametic sex produces
two types of sperms, half with X chromosome and half without X chromosome in equal
proportions. The sex of the offspring depends upon the sperm that fertilizes the egg, each
of which carries a single X chromosome. Thus fertilization between male and female
gametes always produced zygotes with one ‘X’ Chromosome from the female, but only
50% of the zygotes have an additional X Chromosome from the male. In this way, ‘XO’
and ‘XX’ types would be formed in equal proportions, the former being males and the
latter being females.

Parents Female X Male

2A + XX X 2A + XO
A+X A+O
A+X 2A + XX Female 2A + XO Male

ii) XX – XY: In man, other mammals, certain insects including Drosophila, certain
angiospermic plants including Melandrium, the females possess two X chromosomes

3
(XX) and are thus homogametic and homomorphic, while the males possess one X and
one Y chromosome (XY) and are hence heterogametic and heteromorphic. The Y
chromosome is acrocentric not Y shaped as is commonly assumed. When an egg is
fertilized by ‘Y’ bearing sperm, a male is produced.

Parents Female X Male

2A + XX X 2A + XY
Gametes
A+X A+Y
A+X 2A + XX Female 2A + XY Male

b) Heterogametic female: In this mechanism the male possess two homomorphic sex
chromosomes and are thus homogametic, while the female possesses either a single ‘X’
chromosome or one ‘X’ and one ‘Y’ chromosome and are hence heterogametic. To avoid
confusion with earlier types, instead of X and Y, the alphabets Z and W are used. This
mechanism of sex determination is also known as “Abraxas mechanism of sex
determination” (Kuspira and Walker, 1973) The heterogametic females may be of
following two types.

i) ZO – ZZ ii) ZW – ZZ

i) ZO – ZZ: This mechanism is found in certain moths and butterflies. In this case,
female possess one single ‘Z’ chromosome and hence is heterogametic. Male possesses
two Z chromosomes and thus homogametic.

Parents Female X Male

2A + ZO X 2A + ZZ
Gametes
A+Z
A+Z 2A + ZZ Male
A+O 2A + ZO Female
The sex of the offspring depends on the kind of egg.

ii) ZW – ZZ: This system is found in certain insects (gypsy moth) and vertebrates such
as fishes, reptiles and birds. In this system, the female is heterogametic and produces two
types of gametes, one with ’Z’ chromosome and the other with ‘W’ chromosome. On the
other hand, male is homogametic and produces all sperms of same type carrying one ‘Z’

4
chromosome. To prevent confusion with the XX-XY system, the sex chromosomes in
this system are labeled Z and W, but the chromosomes did not resemble Zs and Ws. The
sex of the offspring depends on the kind of egg being fertilized. The ‘Z’ chromosome
bearing eggs produce males, but the ‘W’ chromosome bearing eggs produce females.

Parents Female X Male

2A + ZW X 2A + ZZ
Gametes
A+Z
A+Z 2A + ZZ Male
A+W 2A + ZW Female

II) GENIC BALANCE MECHANISM:

By studying the sex chromosomal mechanism of sex determination, it may appear at first
glance that some genes carried by sex chromosomes i.e. X and Y are entirely responsible
for determining sex. But this may not always be true. Extensive experiments on different
organisms by different workers have revealed the fact that most organisms generally have
inherent potentialities for both sexes and each individual is found to be more or less
intermediate between male and female. Hence may be referred to as inter sex. There
seems to exist a delicate balance of masculine and feminine tendency in the hereditary
compliment of an individual. Such a genic balance mechanism of determination of sex
was first observed and studied by C.B. Bridges in 1921 while working with Drosophila
for the inheritance of vermillion eye colour. “According to this mechanism, the sex of an
individual in Drosophila melanogaster is determined by a balance between the genes for
femaleness located in the X-chromosome and those for maleness located in autosomes.
Hence, the sex of an individual is determined by the ratio of number of its X
chromosomes and that of its autosomal sets, the ‘Y’ chromosome being unimportant. The
ratio is termed as sex index and is expressed as follows.”

Sex index = No. of X chromosomes / No. of autosomal sets = X / A

Different doses of X – Chromosomes and autosome sets and their effect on sex
determination

S. No. Ploidy X-Chromosomes Autosomes Sex index Sex Expression

level Sets of (X/A ratio)
1 Diploid 3(xxx) 2(AA) 1.50 }Super female }

5
 2 Triploid 4(xxxx) 3(AAA) 1.33 or meta female }
3. Haploid 1(x) 1(A) }
4. Diploid 2(xx) 2(AA) }
5. Triploid 3(xxx) 3(AAA) }1.00 Female
6 Tetraploid 4(xxxx) 4(AAAA) }
}
7 Triploid 2(xxy) 3(AAA) 0.67 }
8 Tetraploid 3(xxxy) 4(AAAA) 0.75 } Inter sex
}
9 Diploid 1(xy) 2(AA) } Male
10 Tetraploid 2(xxyy) 4(AAAA) } 0.5
11 Triploid 1(xyy) 3(AAA) 0.33 Super male or meta
male
Individuals with sex index of 0.5 develop into normal males and those with sex index of
1 into normal females. If the sex index is between 0.5 and 1, the resulting individuals will
be neither a female nor a male, but have an intermediate sex expression and is called inter
sex. Such individuals are sterile. Some flies have sex index of >1, such flies have more
pronounced female characteristics than normal females and are called super females or
meta females. These are generally weak, sterile and non-viable. Super male flies have a
sex index value of <0.5 and are also weak, sterile and nonviable.

Bridges drew the observation by crossing triploid females (3A + XXX) with normal
diploid males (2A + XY). From such a cross he obtained normal diploid females, males,
triploid females, intersexes, super males and super females. The occurrence of triploid
intersexes from such a cross clearly established that autosomes also carry genes for sex
determination. Triploid individuals, which had two ‘X’ Chromosomes as in the case of
normal female, here were inter sexes as they had an extra set of autosomes indicating that
the autosomes play a definite role in the determination of sex.

Results obtained from a cross of a triploid (3A+XXX) female fly with a diploid
(2A+XY) male fly in Drosophila

Parents Female X Male

Phenotype Triploid X Diploid
Genotype 3A + XXX X 2A + XY
Gametes
A+X A+Y
2A + XX 3A + XXX Triploid female 3A + XXY Triploid inter sex

6
 A+X 2A + XX Diploid female 2A + XY Diploid male
2A + X 3A + XX Triploid inter sex 3A + XY Super male
A + XX 2A + XXX Super female 2A + XXY Diploid female

III. Male haploidy or haplodiploid mechanism or arrhenotokous parthenogenesis:

It is common in hymenopterous insects (ants, bees, wasps). In honey bees, queens usually
mates only once during its life time and the sex ratio of offspring is under the control of
queen. Fertilized eggs develop into diploid female and those eggs which the queen
chooses not to be fertilized develop parthenogenetically into haploid but fertile males
(drones). This phenomenon is known as arrhenotoky and is a form of reproduction as
well as a means of sex determination. Meiosis is normal during oogenesis in case of
females and produces all haploid eggs. But crossing over and reduction division fails to
occur during spermatogenesis in males due to their haploid nature. Thus arrhenotokous
parthenogenesis determines the sex in hymenopterans and sex chromosomes have no
identity here (unlike Drosophila). It seems that heterozygosity for specific genes induces
femaleness. The haploid can never be heterozygous. Most of the eggs laid in the hive will
be fertilized and developed into worker females. Further during investigation, it has been
found that the quantity and quality of food available to the diploid larvae determines
whether that female will become a sterile worker or a fertile queen. The diploid larva,
which feed on royal jelly, develop into fertile female called queen and the remaining
larvae give rise to workers, which are sterile females. Thus, environment here determines
sterility or fertility but does not alter the genetically determined sex.

IV. SEX DETERMINATION DUE TO ENVIRONMENTAL FACTORS

In many reptiles, sex is determined by environmental factors like temperature. In most

turtle species only females are produced at high temperature (30 – 35ºC) while only
males are produced at low temperatures (23 – 28ºC). Sex ratio changes suddenly from all
males to all females with just change in temperature of 2ºC during the incubation. In
Crocodiles and Lizards, (reverse is the case) the males are produced at high temperature
while females are produced at low temperature. In Bonellia viridis , a marine worm, all
larvae are genetically and cytologically similar. If a particular larva settles near proboscis
of adult female, it becomes a male individual. If larva develops freely in water, it
becomes a female. In some plants, sex determination depends on day length, temperature
and hormones. For example, in cucumber (Cucumis sativa) and muskmelon, treatment
with ethylene enhances production of female flowers.

7
THE ROLE OF SEX CHROMOSOMES

The phenotype associated with sex-chromosome anomalies allow us to make several

inferences about the role of sex chromosomes in human sex determination.

1. The X chromosome contain genetic information essential for both sexes; at least
one copy of an X chromosome is required for human development.
2. The male determine gene is located on the Y chromosome. A single copy of this
chromosome, even in the present of several X chromosomes, produce a male
phenotype.
3. The absent of the Y chromosome results in a female’s phenotype
4. Genes affecting fertility are located in the X and Y chromosomes. A female
usually need at least two copies of the X chromosome to be fertile.
5. Additional copies of the X chromosome may upset normal development in both
males and females, producing physical and mental problems that increases as the
number of extra X chromosome increases

8
 SEX LINKAGE

SEX-LINKED CHARACTERISTICS

The characters for which genes are located on sex or ‘X’ or analogous ‘Z’ chromosomes
are known as sex linked traits. Such genes are called sex linked genes and linkage of such
genes is referred to as sex linkage. Inheritance of such genes or characters is known as
sex linked inheritance. The sex linkage was first discovered by T.H. Morgan in
Drosophila and the first sex linked gene found in Drosophila was recessive gene ‘w’
responsible for white eye colour

Parents Female X Male

Phenotype White eye X Red eye
Genotype Xw Xw X X+Y
Gametes
X+ Y
+ w
Xw XX XwY
Red eye female White eye male
When white eyed females are crossed with wild type (red eye) males, all the male
offspring have white eyes like the mother and all the female offspring have red eyes like
their father. In human beings, colour blindness and haemophilia (bleeder’s disease) are
well known hereditary characters showing a peculiar relationship to sex. Colour
blindness: A marriage between colour blind woman and a normal man gives rise to all
normal dauthers and colour blind sons.

Parents Mother X Father

Phenotype Colour blind X Normal
Genotype Xc Xc X X+Y
Gametes
X+ Y
+ c
Xc XX Xc Y
Normal daughter Colour blind son
The allele for colour blindness (c) is found on both ‘X’ chromosomes of mother and
therefore she is colour blind. The only one ‘X’ chromosome of father in this marriage
carries a wild type (+) allele and hence he has normal vision. The ‘Y’ chromosome lacks
both the alleles (+ and c). The reduction division produces one kind of egg in contrast to
two kinds of sperms. Fertilization results in the usual sex ratio of 1 male : 1 female. All
the daughters have normal vision since they receive dominant allele ‘+’ from their father.
All the sons are colour blind, because their single ‘X’ chromosome derived from mother

9
carries the allele ‘c’ for colour blindness. This result is known as crisscross inheritance
because daughters are normal like father and sons have colour blindness like mother.
However, the daughters are heterozygous carriers. This crisscross method of inheritance
is characteristic of sex-linked genes. This peculiar type of inheritance is due to the fact
that Y chromosome carries no alleles homologous to those on the X chromosome. Thus
males carry only one allele for sex linked traits. This one allelic condition is termed as
hemizygous in contrast to homozygous and heterozygous possibilities in female. The
expression of recessive gene in hemizygous condition is termed as pseudo-dominance.

The inheritance of colour blindness can be studied in the following three other possible
types of marriages:

a) Marriage between normal woman and colour blind man:

Parents Mother X Father

Phenotype Normal X Colour blind
Genotype X+ X+ X Xc Y
Gametes
Xc Y
X+ X+ Xc X+ Y
Normal daughter (Carrier) Normal son

b) Marriage between normal (carrier) woman and normal man:

Parents Mother X Father

Phenotype Normal X Normal
Genotype X+ Xc X X+ Y
Gametes
X+ Y
+
X X+X+ Normal daughter X+Y Normal son
Xc X+Xc Normal daughter XcY Colour blind son

c) Marriage between normal (carrier) woman and colour blind man:

Parents Mother X Father

Phenotype Normal X Colour blind
Genotype X+ Xc X Xc Y
Gametes

10
 Xc Y
+
X X+ Xc X+ Y
Normal daughter Normal son
Xc Xc Xc Xc Y
Colour blind daughter Colour blind son

Results of possible four marriages make it clear why there are more colour blind males
than females in the population. In three marriages colour blind sons were produced where
as in only one of the marriages, colour blind daughters were observed, where the mother
is heterozygous (carrier) and the father is colour blind. Nearly all colour blind women
must come from the last type of marriage, since the only other possible source of colour
blind females is mating between two colour blind persons – naturally a rare occurrence.

Haemophilia is a recessive sex linked disease and the inheritance pattern of haemophilia
is similar to that of colour blindness in human beings. Genes present in the non-
homologous region of the Y chromosome pass directly from male to male. In man, the
genes present on Y chromosome (holandric genes) such as the gene causing
hypertrichosis (causing excessive development of hairs on the pinna of ear) are
transmitted directly from father to son.

SEX INFLUENCED INHERITANCE

Sex influenced genes are the autosomal genes present in both males and females, whose
phenotypic expression is different in different sexes in such a way that they act as
dominant in one sex and recessive in the other i.e. in a pair of alleles one seems to be
dominant in males while the other in females.

Eg.: Pattern baldness in human beings and horns in sheep. Pattern baldness in human
beings is a condition in which a low fringe of hair is present on the head in human beings.
It is a genetically inherited condition, where the allele for baldness B is dominant in
males and recessive in females. In heterozygous condition, males are bald and females
are non bald.

If a woman heterozygous for this gene marries a heterozygous bald man, in the offspring,
the ratio of bald to non-bald in males is 3 : 1, while in females it is 1 : 3.

Inheritance of pattern baldness in human being

Parents Mother X Father

Phenotype Bald X nonbald
11
Genotype b+b+ X bb
Gametes
F1

b+b

Female (Nonbald) Male (Bald)

Phenotypic expression of pattern baldness in man and woman

Genotype Phenotype
Female Male
+ +
b b Nonbald Nonbald
b+b Nonbald Bald
bb Bald Bald

Sex limited characters or Secondary Sexual Characters Sex limited genes are autosomal
genes, whose phenotypic expression is limited to one sex only. Their phenotypic
expression is influenced by the sex hormones. The sex limited genes are mainly
responsible for secondary sex characters in cattle, human beings and fowl. Eg.: milk
production in cattle, beard development in human beings, plumage in male fowls etc.
Milk production in cattle : Just as the cow, the bull carries genes for milk production, but
the bull obviously cannot express this trait. Bull may however transmit these genes for
high milk production to the female progeny and the male progeny are unable to express
this trait. Some bulls are so well andowed with such genes that they are known to breed
calves, which always yield greater milk than their mothers. However, in plants no
secondary sexual characters are known except the absence of one or the other sporangia.
Differences between sex linked and sex limited characters

Sex Linked characters Sex Limited characters

They are located on sex or X chromosome They are located on sex chromosomes or
autosomes

They can express in both the sexes They can express in one sex only

Include characters not related to sex Include primary and secondary sex characters

Examples: White eye in Drosophila, Examples: milk production in cattle, beard

12
 haemophilia and colour blindness in development in human beings, plumage in male
human beings fowls etc

Inter sex

In a few rare cases, various mixtures of male and female characteristics may occur in
animals, which normally have separate sexes because of var ious abnormalities of
chromosomes or hormonal deficiencies. Such individuals are called inter sex. These can
be of two types.

1. Pseudo -hermaphrodites: In mammals there are rare cases in which, both sexes are
well developed in one body, these are abnormal and hence sterile.

2. Gynandromorphs or gynanders: Among animals and insects that do not have sex
hormones, there may be sex intergrades with distinct areas of the body showing male and
female tissues. For example: Drosophila gynander, a bilateral sex mosaic, is male on one
side and female on the other.

13
 DISPUTED PATERNITY

When a child is born he or she carries the DNA of both the parents. Both parents are
equally responsible for the birth of the child. The man who has contributed his sperms to
this child is considered to be the biological father. But there is another term called legal
father, a legal father is the one who is recognized as the father of the child by the law.
The legal father may not be the biological father of that child. The word paternity means
legal fatherhood of the child. Every child has a biological father but not legal father. A
legal father has the right to support the child financially, take care of the child etc.

In many societies it has been found that the child is born from unmarried parents, in this
case this biological father is not considered as the legal father. Even if the name is written
in the birth certificate then also he is not the legal parent of the child. He has to establish
it with the help of law by recognition of parental (ROP) process and by court of law.

Paternity disputes occur when the father claims that he is not the biological father of that
child and refuse to take the responsibility of the child, however the mother claims that the
man is the biological father of her child.

The question of disputed paternity may arise in:

1. Illegitimacy

2. Posthumous birth

3. Suppositious child

4. Nullity of marriage

5. Divorce

6. Inheritance of property

7. Guardianship

8. Maintenance

As we know that only a legitimate child can inherit the property. In case of Lohi and
Radhika Singh the Supreme Courted states that if a man and woman who are unmarried
and stay together for a long time and behave like a married couple then there is
presumption of a valid marriage and the child born will be legitimate and will have the
right to inheritance and succession.

14
Under Section 3(57) of the General Clauses Act, 1897, the definition of son includes an
adopted son and under Section 12 of the Hindu Adoption and Maintenance Act, 1956, an
adopted son or daughter is entitled to succeed to the estate of his adoptive parents.

APPROACHES TO SOLVE DISPUTED PATERNITY:

Disputed paternity can be solved through different approaches:

1. Morphological approach between the child and parent.

2. Serological approach

3. Immunological approach through Human Leukocyte Antigen (HLA) typing

4. DNA typing (DNA Profile) which include Variable number tandem repeats (VNTR),
short tandem repeats (STR), polymerase chain reaction (PCR).

MORPHOLOGICAL APPROACH BETWEEN THE CHILD AND THE PARENT:

Every trait is transferred from parents to children. Each parent contributes equally in the
formation of zygote. So it is normal that the child represents the characters similar to its
biological parents. The dominant characters are expressed in phenotypically. The child
resembles his or her parents many times in features like figure, skin color, nose form,
eyes, stature, hair and other personal details. Many a times some deformaties or
developmental details are also seen in the child. Many diseases are transformed from
father to boy as Y chromosome in inherited from father to son and many diseases are
carried from mother to son as X chromosome is transferred from mother to son. In case
of female child one X chromosome is transferred from mother to child and another X
chromosome from father to child. In female child many times genotype is not represented
phenotypically i.e., morphologically as because they are the carrier. Sometimes child
represents the trait that their grandparents have, as traits are inherited and transferred
from one generation to another. For example: Hypertrichosis pinnae auris is a Y-linked
autosomal dominant trait in which the helix of the ear shows hair growth. This trait is
transformed from father to son as it is located on the Y chromosome.

Few Mendelian traits like ear lobe, tongue rolling etc. also help in disputed paternity.
Joint ear lobe is a dominant trait and attached ear lobe is a recessive trait. Rolling-tongue
is a dominant trait while not rolling-tongue is a recessive trait.

15
 SEROLOGICAL APPROACH

ABO Blood Group System

ABO blood group was discovered by Landsteiner in 1900. ABO blood group has antigen
A and B. Blood group of a person is identified with the antigen and anti-body. A blood
group carry only A antigen, B blood group carry only B antigen, AB blood group carry
both A and B antigen and O blood group carry no antigen. Again, A blood group has
anti-B antibody in the plasma, blood group B has anti-A antibody, blood group O has
anti-AB antibody and blood group AB has no antibody.

BLOOD GROUP ANTIGEN PRESENT ANTIBODY PRESENT

A A anti-B
B B anti-A
AB A,B none
O none anti-A and B
ABO blood group has three allelic genes A, B and O. Genotype and phenotype of
different blood group is given below:

GENOTYPE PHENOTYPE
OO O
AO A
AA A
BO B
BB B
AB AB
The factors that determine the blood group of the child are inherited from parents. But
blood grouping test can never affirmatively fix paternity of a man, but they may
exonerate him. To understand how this blood grouping helps in knowing that a man is not
the father of the given child, it is necessary to understand the way in which blood groups
are inherited: during zygote formation haploid sperm cell and a haploid ovum fuses with
each other if father has blood group AB then two types of sperm will be produced one
that will carry A and the other that will carry B. If the mother also has AB blood group
then she will also produce two types of ovum one that will carry A and the other that will
carry B. The genotype of the child can be AB, AA, BB and AB. So there is 50% chance
of the child to have AB blood group and 25% chance for AA and BB blood group.

If the father has blood group B then the genotype of the father can be BB and BO, if the
father has genotype BB then all the sperms will carry B genes and if the genotype is BO

16
then half sperms will carry B and the other half will carry O genes. Now if the mother
also have B blood group then genotype can be BB and BO, if the genotype is BB then all
the ovum will carry B genes and if the genotype is BO then half of the ovum will be
carrying B genes and the other half will carry O genes. Then the child will have B blood
group but if father and mother have genotype BO and BO then it is possible that the child
will have O blood group. As because the possible genotypes will be BB, BO, BO, and
OO. If both the parents have blood group A then the possible genotype can be AA or AO
and their child will have A blood group. But if the parents have genotype AO and AO
then the child can have blood group A and O.

If one of the parents has blood group AB and other have blood group O than the child can
have blood group A or B.

ABO blood group also helps in establishing non paternity, if the blood group of the father
and mother is known then easily we can get to know the blood group of the child and if
the child does not have the possible blood group then non paternity is established.

PARENT’S BLOOD TYPES POSSIBLE CHILD NOT POSSIBLE CHILD

A and A A,O B, AB
A and B A,B,AB,O no
A and AB A, B, AB O
A and O A,O B, AB
B and B B,O A, AB
B and AB A, B, AB O
B and O B,O A,AB
AB and AB A, B, AB O
AB and O A,B AB,O
O and O O A,B,AB

MN Blood Group System

MN blood group also helps in determining blood group: the MN blood group is
controlled by autosomal loci which are present in the chromosome 4 and two alleles
responsible for this blood group MN are LM and LN. This blood group is determined by
the presence of glycoprotein on the surface of the red blood cell. The three phenotypes
which are possible are MM, NN and MN. This is also inherited from one generation to
another so it helps in determining the paternity.

17
If the mother has blood group M, then the genotype is MM and the child has blood group
MN then the father should have blood group N and the genotype is NN. if the mother has
blood group M, then the genotype is MM and the child is of blood group M, genotype is
MM then the blood group of the father of the child is also M or MN. If the blood group of
the mother is N and the child also have the blood group N then the blood group of the
father is also N or MN.

Immunological Approach through Human Leucocyte Antigen Typing:

This is also a kind of serological test which helps in determining paternity. Human
Leukocyte was developed by Dr. Paul I. Terasaki, Professor of Surgery at the University
of California at Los Angeles, in 1964. He discovered this system to minimize the
possibility of rejection of transplant organ.

The Human Leukocyte Antigen system is based upon the identification of antigens,
substances that stimulate antibody production when introduced into another human body.
Because the HL-A test detects antigens by using antisera (antibodies), it is known as a
serologic test (Sterlek, 1980). As genes are inherited from the parents, by making
calculations on the basis of the antigens present on the surface of the white blood cells
probability of paternity in calculated. This test is based on one of the two ways:

That there may be exclusion; the man is not the father of the child according to the
principles of genetics.

Another, there may be inclusion; the man can be the father of the child or may have
similar genetic makeup.

The advantage of HLA over blood typing is that all HL-A types are relatively rare. If the
accused man shares high percentage of combination of HL-A types than the man could be
the father of the child. But high rate of exclusion are possible products of multiple
testing, costs and diminishing returns render excessive multiple testing impractical
(Sterlek, 1980).

DNA Typing:

DNA is the deoxyribonucleic acid which is the genetic material and inherited from both
the parents. DNA is the ideal source for identification as it is unique in every individual,
it shows the blue print. It is a method for identification of the individual used in forensic
science. It also helps in the parental analysis by taking the allele size for all microsatellite
markers. This technique was developed by Alec Jeffreys in 1987.

18
Techniques which are involved are:

a. Variable Number Tandem Repeats (VNTR)

These are short tandem sequences of 10-100 base pairs which are repeated. These regions
vary from individual to individual in numbers. Each variant act as an inherited allele
allowing them to use as personal identification and parental identification. It is almost
impossible having equal VNTR regions in two unrelated person.

Multilocus DNA fingerprinting is one of the important tools in determining paternity

because they provide great information, somatic stability and Mendelian inheritance.

b. Short Tandem Repeats (STR)

These are short arrays of tandem repeated sequences of 2-6bp in length. These are the
polymorphic regions which vary from individual to individual.

These are widely used in forensics since 1993. STR is dependent on the PCR which
confers much greater sensitivity on the test system.

This technique does not consume much time and is used in forensic case work. Greater
sensitivity allows the use of more convenient sample.

The relative reduction in discriminating power with respect to SLP profiling has a more
profound effect in parentage testing, where generally only one allele at each locus is
informative, than in identity testing, where a match at both alleles is required. This
distinction means that a six locus STR system such as the Second Generation Multiplex
(SGM) has been designed which has an equivalent discriminating power in identity cases
to four SLPs but many more STRs are required to provide equivalent paternity indices to
the six SLPs currently used by the UDL laboratory in cases of disputed paternity
(Thomson, 1999).

c. Polymerase Chain Reaction

It was discovered by Kary Mulluis in 1983. It was established as a standard method for
paternity testing. PCR helps in multiplication of a specific region of DNA strand many
times. It is the most accurate and fastest method for determining paternity as DNA is
inherited from parents to the child. Through PCR one can determine the closeness
between two individual or how closely they both are related with each other. The two
DNA source is multiplied and seen whether one is derived from the other or if the two
had similar parentage.

19
A child inherits a unique combination of DNA from its parents. Because scientists have
extensively used PCR for DNA testing, a greater amount of information has been
accumulated to form a database for accurate DNA analysis. This large database enables
paternity testing via PCR to have the highest power of exclusion (Khanagwal, 2012).

According to Chakraborty three human microsatellites which are commonly typed by

PCR are apoB, D17S5 which are also known as D17S30 and D1S80. These markers can
amplify easily and detected after electrophoresis.

Earlier restriction fragment length polymerase (RFLP) is used in determining paternity

but nowadays it is not used much as it requires large amount of DNA and takes a long
time to process. Now new methods like PCR are used.

20
 GENETIC VARIATION

Variations are the small differences that exist between individuals. It can be described as
being either discontinuous or continuous.

These are characters or traits that can be found amongst various species of organisms.
These variations can either heritable or non-heritable.

The heritable characters or variations are those that can be passed though the germ line
(sperms or eggs) during the process of sexual reproduction or through attributes passed
down through the soma such as leaves, stems and roots that occasional serve as
reproductive unit (sexual reproduction).

In sexual reproducing organism, the sperm, pollen and eggs constitute the germ line
containing the attributes in parents (biological variations) which are subsequently passed
on to the offspring. Thus, the germ lines are the bearers of information that controls the
development of the offspring in the direction of the information content of the zygote
which results from the union of the sperm and egg. Developmental errors that do not
involve the germ line may be manifested in the parents, but are not, in general
transmissible to the offspring. Such errors and post-natal accidents are referred to as non-
heritable characters e.g. amputation of limbs, or blindness that results from accidents,
weight lifting, a blacksmith or professional skills. A weight lifter or a blacksmith
develop extra muscles nut this muscle development cannot be passed on to the offspring
unless the offspring is subjected to the same physical exercise as the parents. It is to be
however noted that the environment does contribute to observable biological variations.
Culture traits such as language and mannerisms are passes on from parents to offspring,
but unlike heritable traits, cultural traits depend on cultural environment rather than
parentage. Thus, cultural are also environmentally induced and are mot transmitted
through the germ line. Some traits are partly heritage and partly cultural e.g. body
movement of a woman is usually different from that of a man. Part of the difference is
due to the muscle structure of the female and the distribution of sexual organs such as
breasts and ovary in the females, but a large part of the seductive movements is learnt
from other females.

1. Heritage variations may either be continuous (indiscrete) or discontinuous (discrete).

a. CONTINUOUS OR INDISCRETE VARIATION

21
These are variations that have intermediary values, which are not exact, i.e. indiscrete.
Such variations are mainly morphological or physical variations. Examples are height,
weight, length, skin pigmentation (colour), colour of eyes, fingerprints, etc.

In continuous variation there is a complete range of measurements from one extreme to

the other. Height is an example of continuous variation – individuals can have a complete
range of heights, for example, 1.7, 1.61, 1.62, 1.625, etc metres high. Other examples of
continuous variation include:

Weight, Hand span, Shoe size, Milk yield in cows. Continuous variation is the combined
effect of many genes (known as polygenic inheritance) and is often significantly affected
by environmental influence. Milk yield in cows, for example, is determined not only by
their genetic make-up but is also significantly affected by environmental factors such as
pasture quality and diet, weather, and the comfort of their surroundings.

b. DISCONTINUOUS OR DISCRETE VARIATION

these are variations that do not have intermediary values. That is, the values are exact or
discrete. Such variations are mainly physiological variations. Examples are sickling
character or trait, blood group, Gender, tasters, tongue rolling, haemophilia etc. this is
where individuals all into a number of distinct classes or categories, and is based on
features that cannot be measured across a complete range. You either have the
characteristics or you don’t. bloodgroup are a good example: you are either one blood
group or another you can’t be in between. Such data is called discrete (or categories)
data.

Discontinuous variation is controlled b alleles of a single gene or a small number of

genes. The environment has little effect on this type of variation.

CAUSES OF VARIATION AND ROLE OF MEIOSIS IN CAUSING VARIATION

Variation in the phenotype is caused either by the environment, by genetics, or by a

combination of the two. Meiosis and sexual reproduction introduces variation through
independent assortment of the parental chromosomes; through crossing-over during
prophase I; and through the ransom fertilization that forms the zygote.

Meiosis is a major source of genetic variation within a population. Though chromosome

sets of the four resulting haploid cells look identical, they may be quite different
genetically.

22
Homologous chromosomes of the original diploid cell may have subtle mutational
differences between their genes, and each pair of chromosomes separates independently
during Anaphase, different haploid cells may have quite different combinations of genes
derived from the two parents of this haploid organism.

23
 MUTATION

This is a sudden, random alteration in the genotype of an organism, involving qualitative

or quantitative alterations in the genetic materials itself. It may involve a single gene
(point mutation) or a whole chromosomal aberrations).

Note: changes within genes are called point mutation,

Mutation that span more than one gene are called chromosomal mutations because they
affect the structure, function and inheritance of whole DNA molecules

Mutation may either affect the gene number or chromosome structure.

In biology, mutations are changes to the nucleotide sequence of the genetic material of an
organism. Mutations can be caused by copying errors in the genetic material during cell
division, by exposure to ultraviolet or ionizing radiation, chemical mutagens, or viruses,
or can be induced by the organism, itself, by cellular processes such as hyper mutation. In
muticellular organisms with dedicated reproductive cells, mutations can be subdivided
into germ line mutations, which can be passed on to descendants through the
reproductive cells, and somatic mutations, which involve cells outside the dedicated
reproductive group and which are not transmitted to descendants, usually. If the organism
can reproduce asexually through mechanisms such as cuttings or budding the distinction
can become blurred. For example, plants can sometimes transmit somatic mutations to
their descendants asexually or sexually where flower buds develop in somatically
mutated parts of plants. A new mutation that was not inherited from either plant is called
a de novo mutation. The source of the mutation is unrelated to the consequence, although
the consequence are related to which cells are affected.

Neutral mutations are defined as mutations whose effects do not influence the fitness of
an individual. These can accumulate over time due to genetic drift. It is believed that the
overwhelming majority of mutations have no significant effect on an organism’s fitness.
Also DNA repair mechanisms are able to mend most changes before they become
permanent mutations, and many organisms have mechanisms for eliminating otherwise
permanently mutated somatic cells.

Gene mutation in man

Base deletion means a single base is omitted. This will affect all subsequent amino acids
in that protein and the effects are therefore likely to be sever. Base substitution means
that one base is replaced by another. This will affect a single amino acid, and normally
this has little effect unless the active site of an enzyme is affected. One example where

24
the effects are dramatic is sickle-cell anaemia, which is caused by a single amino-acid
substitution occasionally mistakes occur during DNA replication and protein synthesis.
Any alteration in the structure of a gene results in a mutation. Mutation occurs during
DNA replication when the chemical structure of gene undergoes random modifications.
Once a change has occurred, the altered genes continue to replicate in their changed form
unless another mutation occurs. Sometimes mutations may occur in any living cell, they
are most important when they occur in gametes because then the change affects the traits
of following generations. Most mutations harm an organism. If a mutation occurs in a
gene sequence that codes for a particular protein, the mutation may result in a change in
the amino acid sequence directed by the gene. This change, in turn, may affect the
function of the protein. The implications can be significant: the amino acid sequence
distinguishing normal hemoglobin from the altered form of hemoglobin responsible for
sickle-cell anaemia differs by a single amino acid.

Some mutations may be neutral or silent and do not affect the function of a protein.
Occasionally a mutation benefits an organism. Over the course of evolutionary time,
however, mutations serve the crucial role of providing organisms with previously
nonexistent proteins. In this way mutations are a driving force behind genetic diversity
and the rising of new or more competitive species better able to adapt to changes, such as
climate variations, depletion of food sources, or the emergence of new types of disease.

Mutations can produce a change in any region of a DNA molecule. In a point mutation,
for example, a single nucleotide replace another nucleotide. Although a point mutation
produces a small change to the DNA sequence, it may cause a change in the amino acid
sequence, and thus the function of a protein. For more serious are mutations that involve
the addition or deletion of one or more bases from a DNA molecule. Adding or
subtracting even a single base from a normal sequence during transcription can disrupt
translation by shifting the “reading frame” of every subsequent codon. For example, am
mRNA strand may include two codons in the following sequence: AUG UGA. The
addition of a cytosine base at the beginning of this sequence shifts the “spelling” of these
codons so that they read: CAU GUG. This may result in an incorrect amino acid
sequence during translation, or the protein may be truncated. Known as frameshift
mutation, this type of alteration could result in the production of a protein with no real
function or one with a harmful effect. Sometimes mutations are caused by transposition,
in which long stretches of DNA (containing one or more genes) move from one
chromosome to another. These jumping genes, called transponsons, can disrupt
transcription and change the type of amino acids inserted into a protein.

25
Transponsons rearrange and interrupt genes in a way that generally improves the genetic
variation of a variation of a species.

While mutations can occur spontaneously, some can be caused by exposure ti physical or
chemical agents in the environment call mutagens. Common environmental mutagens
include ultraviolet rays from the sun and various chemicals, such as asbestors, cigrarette
smoke, and nitrous acid. High energy radiation, such as medical X rays, can cause DNA
strands to break, leading to the deletion of potentially important genetic information.
Radiation damage can also affect an entire chromosome, disrupting the function of many
genes.

In chromosomal translocation, a piece of one chromosome breaks off and merges with
another chromosome. In some cases, large sections of chromosomes may break off and
be lost. The cell has highly effective self-repair mechanisms that can correct the harmful
changes made by mutations and prevent some mutations from being passed on. Some 50
specialized enzymes located different types of faulty sequences in the DNA and clip out
those flaws.

Another repair mechanism scans DNA after replication and marks mismatched base pairs
for repair.

MUTATIONS: TYPES AND CAUSES

The development and function of an organism is in large part controlled by genes.

Mutations can lead to changes the structure of an encoded protein or a decrease or

complete loss in its expression. Because a change in the DNA sequence affects all copies
of the encoded protein, mutations can be particularly damaging to a cell or organism. In
contrast, any alteration in the sequence of RNA or protein molecules that occur during
their synthesis are less serious because many copies of each RNA and protein are
synthesized.

Geneticists often distinguish between the genotype ad phenotype of an organism. Strictly

speaking, the entire set of genes carried by an individual is its genotype, whereas the
function and physical appearance of an individual is referred to as its phenotype.
However the two terms commonly are used in a more restricted sense: genotyprd usually
denote whether an individual carries mutstion in a single gene (or a small number of
genes), and phenotype denotes the physical and function consequences of that genotypr.

MUTATIONS ARE RECESSIVE OR DOMINANT

26
A functional genetic difference between organisms is whether their cells carry a single set
of chromosome or two copies of each chromosome. The former are referred to as haploid
the latter, as diploid. Many simple unicellular organisms are haploid, whereas complex
multicellular organism (e.g., fruit flies, mice, humans) are diploid.

Different forms of a gene (e.g., normal and mutant) are referred to as alleles. Since
diploid organism carry two copies of each gene, they may carry identical alleles, that is,
be homozygous for a gene, or carry different alleles, that is, be heterozygous for a gene.
A recessive mutation is one in which both alleles must be mutant in order for the mutant
phenotype to be observe; that is, the individual must be homozygous for the mutant allele
to show the mutant phenotype.

In contrast, the phenotype consequences of a dominant mutation are observed in a

heterozygous individual carrying one mutant and one normal allele Recessive mutations
inactivate the affected gene and lead to a loss of function, for instance, recessive
mutations may remove part of or all the gene from the chromosome, disrupt expression of
the gene, or alter the structure of the encoded protein, thereby altering its function.

Conversely, dominant mutations often lead to a gain of function. For example, dominant
mutations may increase the activity of a given gene product, confer a new activity on the
gene product or lead to its inappropriate spatial and temporal expression. Dominant
mutations, however, may be associated with a loss of function. In some cases, two copies
of a gene are required for normal function, so that removing a single copy leads to mutant
phenotype. Such genes are referred to as a haplo-insufficient. In other cases, mutations in
one allele may lead to a structural change in the protein that interferes with the function
of the wild type protein encoded by the other allele. These are referred to as dominant
negative mutations. Some alleles can be associated with both a recessive and a dominant
phenotype. For instance, fruit flies heterozygous for the mutant stubble (5b) allele have
short and study body hairs rather than the normal long, slender hairs; the mutant allele is
dominant in this case. In contrast, flies homozygous for this allele die during
development. Thus the recessive phenotype associated with this allele is lethal, whereas
the dominant phenotype is not.

Inheritance patterns of recessive and dominant mutation differ

Rescessive and dorminant mutations can be distinguished because they exhibit different
patterns of inheritance. To understand why, we need to review the type of cell division
that give rise to gametes (sperm and egg cells in higher plants and animals). The body
(somatic) cells of most multicellular organisms divide by mitosis whereas the germ cells

27
that give rise to gametes undergo meiosis. Like body cells, premeiotic germ cells are
diploid, containing two of each morphologic type of chromosome. Because the two
members of each such pair of homologous chromosomes are descended from different
parents, their genes are similar but not usually identical. Single-celled organisms (e.g.,
the yeast S. cerevisiae) that are diploid at some phase of their life cycle also undergo
meiosis.

One round of DNA replication, which makes the cell 4n, is followed by two separate cell
divisions, yielding four haploid (1n) cells that contain only one chromosome of each
homologous pair. The apportionment, or segregation, of homologous chromosomes to
daughter cells during the first meiotic division is random; that is, the maternally and
paternally derived member of each pair. Called homologous, segregate independently,
yielding germ cells with different mixes of paternal and maternal chromosomes. Thus
parental characteristics reasserted randomly into each new germ cell during meiosis. The
number of possible varieties of meiotic segregants is 2n, where n is the haploid number of
chromosomes. In the case of a single chromosome, meiosis gives rise to two types of
gametes; one type carries the maternal homolog and the other carries the paternal
homolog.

Now let’s see what phenotypes are generated by mating of wild-type individuals with
mutants carrying either a dominant or a recessive mutation. Half the gametes from an
individual heterozygous for a dominant mutation in a particular gene will have the wild-
type allele, and half will have the mutant allele. Since fertilization of female gamete by
male gametes occur randomly, half the first filal (F1) progeny resulting from the cross
between a normal wild-type individual and a mutant individual carrying a single
dominant allele will exhibit a mu-tant phenotype. In contrast, all the gametes produced by
a mutant homozygous for a recessive mutation will carry the mutant allele. Thus, in a
cross between a normal individual and one who is homozygous for a rescessive mutation,
more of the F1 progeny will exhibit the mutant phenotype. However, one fourth of the
progeny from parents both heterozygous for a recessive mutation will show the mutant
phenotype.

Mutations involve large or small DNA alteration

A mutation involving a change in a single base pair, often called a point mutation, or a
deletion of a few base pairs generally affect the function of a single gene. Chamge in the
single base pair may produce one of the three mutation:

28
 - Missense mutation, in which a stop codon replace in which one amino acid
codom, leading to premature termination of translation
- Frame shift mutation, which causes a change in the leading frame, leading to
introduction of unrelated amino acid into the protein, generally followed by a stop
codon small deletions have effects similar to those of frame shift mutations,
although one third of these will be in frame and result in removal of small amount
of contiguous amino acid.

The second type of mutation involves large scale changes in chromosome structure and
can affect the functioning of numerous genes, resulting in major phenotypic
consequences. Such chromosomal mutation (or abnormalities) can onvolve deletion or
insertion of several comtiguous genes, inversion or genes on a chromosome, or the
exchange of large segments of DNA between nonhomologous chromosomes,

Mutations arise spontaneously and can be induced

Mutations arise spontaneously at low frequency owing to the chemical instability of

purine and pyrimidine bases and to error during DNA replication. Natural exposure of an
organism to certain environment factors, such as ultraviolet light and chemical
carcinogens (e,g,, aflatoxin B1), also can cause mutations.

A common cause of spontaneous point mutation is the deamination of cytosine to uracil

in the DNA double helix. Subsequent replication leads to a mutant daughter cell in which
a T-A base pair replace the wild-type C-G base pair. Another cause of spontaneous
mutations is coping errors during DNA replication. Although replication generally is
carried out during high fidelity, errors occasionally occur. In order to increase the
frequency of mutation in experimental organisms. Researchers often treat them with high
doses of chemical mutagens or expose them to ionizing radiation; mutations arising in
response to such treatment are referred to as induced mutants. Generally, chemical
mutagens induce point mutations, whereas ionizing radiations give rise to large
chromosomal abnormalities.

Ethyl methane sulfonate (EMS), a commonly used mutations, alkylates guanine in DNA
forming 06-ethyguanine. During subsequent DNA replication, 06-ethyguanine direct
incorporation of deoxythymidylate, not deoxycytidylates, resulting in formation of
mutant cells in which a G-C base pair is replace with an A-T base pair.

Some human disease are caused by spontaneous mutations

29
Many common human diseases, often devastating in their effects are due to mutations in
single genes. Genetic diseases arise by spontaneous mutations in germ cells (egg and
sperm) which are transmitted to future generations, for example sickle cell anemia, which
affects in 500 individuals of Africa descent, is caused by a single missense mutation at
codon 6 of the βglobin gene; as a result of this mutation, the glutamic acid at position 6 in
the normal protein is change to a valine in the mutant protein. This alteration have a
profound effect on hemoglobin, the oxygen-carrier protein of erythrocytes. Which consist
of two α-globin and two β-globin subunits. The deoxygenated form of the mutant protein
is insoluble in erythrocytes and form crystalline arrays. The erythrocytes of effected
individuals become rigid and their transit through capillaries is blocked, causing severe
pain and tissues damage. Because the erythrocytes of the heterozygous individuals are
resistant to the parasite causing malaria, which is epidemic in Africa, the mutant allele
has been maintained. It is not that individual of African descent are more likely than
others to acquire a mutation causing the sickle-cell defect, but rather te mutation has been
maintained in this population by interbreeding.

Spontaneous mutation in somatic cells (i.e. non-germline body cells) also is an important
mechanism in certain human diseases, including retinoblastoma, which is associated with
retinal tumors in children (see. The hereditary form of retinoblastoma, for example,
results from a germline mutation is one Rb allele and a second somatically occurring
mutation in the other Rb allele.

When an Rb heterozygous retinal cell undergoes somatic mutation, it is left with no

normal allele as a result; the cell proliferates in an uncontrolled manner, giving rise to a
retinal tumor. A second form of this disease, called sporadic retinoblastoma, results from
two independent mutations disrupting both Rb allele since only one somatic mutation is
required for tumor development in children with hereditary retinoblastoma. It occurs at a
much higher frequency than the sporadic form, which requires acquisition of two
independently occurring somatic mutations. The Rb protein has been show to play a
critical role in controlling cell division).

SUMMARY

- Diploid organism carry two copies (alleles) of each gene, where are haploid organism
carry only one

- Mutations are alterations in DNA sequences that result in changes in the structure of a
gene. Both small and large DNA alteration can occur spontaneously. Treatment with
ionizing radiation or various chemical agents increases th frequency of mutations

30
- recessive mutations lead to a loss of function, which is masked if a norma copy of the
gene is present. For the mutant phenotype to occur, both alleles must carry the mutation.

- dominant mutations lead to a mutant phenotype in the presence of a normal copy of the
gene.

The phenotypes associated with dominant mutations may represent either a loss or a gain
of function.

- In meiosis, a diploid cell undergoes one DNA replication and two cell divisions,
yielding four haploid cells. The numbers of each pair of homologous chromosomes
segregate independently during meiosis, leading to the radoms rassortment of material
alleles in the gametes.

- dominate and recessive mutations exhibit characteristics segregation patterns in genetic

crosses. Two classes of mutations are spontaneous mutations (molecular decay) and
induced mutations caused by mutagens

INDUCED MUTATIONS

Mutagen or mutagenic agent

This is any agent that brings about mutation. It may be physical or chemical.

a. Physical mutagenic agemts

These include (i) temperature and (2) radiations. The radiation may be ionized e,g X-
rays and cosmic rays or non-ionizing, e.g. ultraviolet (UV) rays.

b. Chemical mutagenic agents

i. Reactants – these reacts with the purine and pyrimidine bases, e.g. nitrous
acid, formaldehyde.
ii. Base analogues – these reacts with the pyrimidine or purine base, e.g. 5-
bromouracil (a pyrimidine analogue), 2-amino-purine analogue).
iii. Acridine dye e.g acridine orange, acridine yellow and proflavin
iv. Alkylating agents e,g mustard gases
v. Carcinogen e.g methyl cholanthrene
vi. Acids e.g phenols

GENETIC SYNDROMES

Down Syndrome

31
This was formerly called Mongolism. It is a syndrome as it consists of a set of
abnormalities. It occurs as a result of nondisjunction (failure of human chromosomes to
separate during meiotic anaphase. People with down syndrome have subnormal
intelligence and a characteristics facial appearance, including a folding of the eyelid
reminiscent of the epicanthic fold of Asiatic people. A karyotype of the chromosome 21,
so the condition is also know as trisimy 21.

Down syndrome children have slight to severe mental retardation.

Turner Syndrome

Person who have turner syndrome are females; they do not undergo puberty and their
female secondary sex characteristics remain immature; menstruation is usually absent,
breast development is slight, and the pubic hair is sparse. This syndrome is seen in 1 of
300 female births.

Affected women are usually short and have a low hairline, a relatively broad chest with
wide space nipples, and folds of skin on the neck. Their intelligence is usually normal.
Most women who have turner syndrome are sterile.

Klinefelter Syndrome

Person who have Klinefelter syndrome, which occur with a frequency of about 1 in 1000
male births, have cells with one or more Y chromosomes. The cells of most males having
this condition are XXY, but cells of few Klinefelter males are XXXY, XXXXY, or
XXYY. Person with this condition, though male, frequently have small testes, some
breast enlargement, and reduced facial and public hair. They are often taller than normal
and sterile; most have normal intelligence.

32
BIOTECHNOLOGY AND NUCLEIC ACIDS

BIOTECHNOLOGY

Biotechnology, the manipulation of biological organisms to make products that benefit

human beings. Biotechnology contributes to such diverse areas as food production, waste
disposal, mining, and medicine.

Although biotechnology has existed since ancient times, some of its most dramatic
advances have come in more recent years. Modern achievements include the transferal of
a specific gene from one organism to another (by means if a set if genetic engineering
techniques know as transgenics); the maintenance and growth of genetically uniform
plant and animal cell cultures called clones; and the fusing of different types of cells to
produce beneficial medical products such as monoclonal antibodies, which are designed
to attack a specific type of foreign substance.

The first achievement in biotechnology were in food production, occurring about

500oBC. Diverse starins of plants or animals were hybridized (crossed) to produce
greater genetic variety. The offspring from these crosses were then selectively bred to
produce the greatest number of desirable traits. Repeasted cyscles of selective breeding
produced many present-day food staples. This method continues ti be used in food
production programs.

The method era of biotechnology had its origin in 1953 when American biochemist
James Watson and British biophysicist Francis Crick presented their double-helix model
of DNA. This was followed by Swiss microbiologist Werner Arber’s discovery in the
1960s of special enzymes, called restriction enzymes, in bacteria. These enzymes cut the
DNA strands of any organism at precise points. In 1973 America geneticist Stanley
Cohen and American biochemist Herbert Boyer removed specific gene from one
bacterium and inserted it into another using restriction enzymes. This event marked the
beginning of recombinant DNA technology, commomly called genetic engineering. In
1977 genes from other organisms were transferred to bacteria. This achievement
eventually led to the first transfer of a human hormones, they produced it along with that
own normal chemical compounds.

In the 1960s an important project used hybridization followed by selective breeding to

increase food production and quality of wheat and rice crops. American agriculturalist
Norman Borlaug who spearheaded the program, was award the Noble peace prize in 1970
in recognition of the important contribution that increasing the world’s food supply
makes to the cause of peace.

33
34
NUCLEIC ACIDS

Nucleic acids, extremely complex molecules produced by living cells and viruses. Their
name comes from their initial isolation from the nuclei of living cells. Certain nucleic
acids, however, are found not in the cell nucleus but in the cell cytoplasm. Nucleic acids
(DNA and RNA) perform a variety of crucial functions in organisms. DNA stores and
transfers genetic information, it serves as the template for the synthesis of new DNA and
RNAs, while RNAs carry out protein synthesis. Nucleic acids contain only a few
different components, but they have great structural diversity. This diversity results from
the many possible combinations of those few components due to the large sizes of DNA
and RNA.

CLASS OF NUCLEIC ACIDS

The two classes of nucleic acids are the deoxyribonucleic acids (DNA) and the
ribonucleic acids (RNA). The backbone of both DNA and RNA molecules are shaped
like helical strands. Their molecular weights are in millions. To the backbones are
connected a great number of small molecules (side group) of four types. the sequence of
these molecules on the strand determines the code of the particular nucleic acid. This
code in turn, signals the cell how to reproduce either a duplicate of itself or the protein it
requires for survival.

DEOXYRIBONUCLEIC ACID (DNA)

Deoxyribonucleic acid (DNA) is the material of which genes are made. This had not been
widely accepted until 1953 when J.D. Watson and F.H, Crick proposed a structure for
DNA which accounted for its ability to self-replicate and to direct the synthesis of
proteins. All living cells (both prokaryotic and eukaryotic) contain double stranded DNA
as their genetic material. The cells of bacteria may have but one strand of DNA, but such
a strand contains all the information needed by the cell in order to reproduces and
identical offspring. The cells of animals contain scores of DNA strand grouped together
in chromosomes. In short, the structure of a DNA molecule or combination of DNA
molecules determines the shape, form, and function of the offspring.

DNA is composed of a series of polymerized nucleotides, joined by phosphodiester

bonds between the 5' and 3' carbons of deoxyribose units. DNA forms a double helix
with these strands, running in opposite orientations with respect to the 3' and 5' hydrozxy
groups.

35
The double helix structure is stabilized by base pairing between the nucleotides, with
adenine and thymine forming two hydrogen bonds, and cytosine and guanine forming
three.

Base + Sugar = nucleoside

Base + Sugar + Phosphate Group = nucleotide

Attached to each sugar residue is one of the four essentially planar nitrogenic organic
bases: Adenine A, Cytosine C, Guanine G, Thymine T,

The two strands coil about each other so that all the bases project inward towards the
helix axis. The two strands are held together by hydrogen bonds linking each base
projecting from one backbone to its complementary base projecting from another
backbone. The base A always binds to T and C always binds to G. This complementary
pairing allows DNA to serve as a template for its own replication.

Adenine A, Cytosine C,

Guanine G, Thymine T

RIBONUCLEIC ACID (RNA)

RNA is similar to DNA but differs in several respects.

1. It is shorter

36
2. It is single stranded (with few exception: few virus)

3. It is nuclear and cytoplasmic

4. It has ribose

5. It has uracil rather than thymine. The other bases are the same.

THERE ARE THREE BASIC TYPES OF RNA:

1. Messenger RNA (mRNA): relatively long strands that encode the information from a
single gene (DNA). It is the template for protein synthesis. This is the product of
transcription. An mRNA is an RNA that is translated into protein. mRNAs are very short-
lived compared to DNA.

► In prokaryotic cells a primary transcript is used directly as an mRNA (often times

before it is even completely transcribed).

► In eukaryotic cells a primary transcript is processed before being exported from the
nucleus as an mRNA:

• A 5'CAP of 7-methyl guanosine is added.

• A poly (A) tail is added to the 3' end of the transcript.

• Introns (intervening sequences) must be cut from the transcript by a process known as
RNA splicing

2. ribosomal RNA (rRNA): Ribosomes are composed of rRNA and protein. The rRNA
forms base pairs with the nucleotides of mRNA during translation (protein synthesis).

3. transfer RNA (tRNA): short (90 nucleotides) RNA molecules responsible for
translating nucleic acid language to protein language. In other words, the "adapter"
molecule that converts nucleic acid sequence to protein sequence.

8.3. Chemical differences between DNA & RNA

► Both RNA and DNA are composed of repeated units. The repeating units of RNA are
ribonucleotide monophosphates and of DNA are 2'deoxyribonucleotide monophosphates.

► Both RNA and DNA form long, unbranched polynucleotide chains in which different
purine or pyrimidine bases are joined by N-glycosidic bonds to a repeating sugar-
phosphate backbone.

37
► The chains have a polarity. The sequence of a nucleic acid is customarily read from 5'
to 3'. For example the sequence of the RNA molecule is AUGC and of the DNA
molecule is ATGC

► The base sequence carries the information, i.e. the sequence ATGC has different
information that AGCT even though the same bases are involved.

38
 GENETIC ENGINEERING

Genetic engineering enables scientists to produce clones of cells or organisms that

contain the same genes.

1. scientists use restriction enzymes to isolate a segment of deoxyribonucleic acid (DNA)

that contains a gene of interest for example, the gene regulating insulin production.

2. a plasmid removed from bacterium and treated wit the same restriction enzyme binds
with the DNA

USES OF GENETIC ENGINEERING

The purposes of doing genetic engineering are many and various. A range of them are
listed below. These include:

1. To repair a genetic "defect" (as with the current early trials of gene therapy in
humans),
2. To enhance an effect already natural to that organism (e.g. to increase its growth
rate),
3. To increase resistance to disease or external damage (e.g. crops - blight, cold or
drought),
4. To enable it to do something it would not normally do: • e.g. getting a micro-
organism to produce human insulin for diabetics, or a sheep to produce a human
blood-clotting protein in her milk, in both cases a transgenic method, • e.g. getting
a tomato to ripen without going squashy - this can be done simply by taking one of
its own genes, turning its "pattern" upside down and putting it back again!

BASIC TOOLS OF GENETIC ENGINEERING

The basic tools of genetic engineering are

1. Cloning vectors: - Which can be used to deliver the DNA sequences in to receptive
bacteria and amplify the desired sequence;

2. Restriction enzymes: Restriction enzymes recognize a specific palindromic sequence

of make a staggered cut, which generates sticky ends, or blund cut, which generate blunt
ends. Which are used to cleave DNA reproducibly at defined sequences and

3. DNA ligases Ligation of the vector with the DNA fragments generates a molecule
capable of replicating the inserted sequence called recombinant DNA.

39
The total No of recombinant vectors obtained when cloning chromosomal DNA is known
as genomic library between there should be at least one representative of each gene in the
library.

Genetic engineering represents a number of methods employed to

i. manipulate DNA outside of cells

ii. place manipulated DNA back into cells
iii. manipulate that DNA following its incorporation back into cells

DNA MANIPULATION

DNA MANIPULATION OUTSIDE OF CELLS

The key to manipulating DNA outside of cells is the existence of enzymes known as
restriction endonucleases. Restriction endonucleases cut DNA only at specific nucleotide
sequences and thus are tools by which DNA may be cut at specific locations. Thus, a
specific gene may be cut out of an organism's genome. Further techniques allow one to
specifically change the nucleotide sequence of the isolated gene.

1. The restriction part of the name derives from the actual use of these enzymes by the
bacteria that make them: restricting the replication of bacteriophages (by chewing up the
bacteriophage DNA)

2. The nuclease part of the name means these enzymes cut DNA

3. The endo part of the name means that they cut DNA in the middle of double helix
strands (rather than chewing DNA up from the ends, i.e., as do exonucleases)

DNA transfer to recipient cell (vector)

• To transfer manipulated DNA back into a cell, one typically first inserts the DNA into a
vector • A vector may be a plasmid (transformation) or a bacteriophage chromosome
(transduction) or both • The vector or plasmid are opened up (cut) using specific
restriction endonucleases • The isolated gene is then inserted into this opening • An
additional enzyme, DNA ligase, then covalently attaches the gene into the vector, thus
making gene and vector into one double helix • The vector may then be transduced or
transformed into a recipient cell • Within that cell the vector is allowed to replicate •
Often these vectors also contain antibiotic-resistance genes which, in the presence of the
appropriate antibiotic, allow only those cells that have successfully received the vector to
replicate

40
Application of recombinant DNA technology

Recombinant DNA technology has been a boom for biological research, allowing
investigator to answer questions about molecular evolution, probe details of gene
organization and control, produce and catalog proteins of interest, and map eukaryotic
genes.

The human genome project involves linkage, physical mapping, and sequencing of the
human genome as well as similar analysis of the genome of some sample species.

Restriction fragment length polymorp

Application of biotechnology in development

Today biotechnology is applied in various fields, in waste management, for example,

biotechnology is used to create new biodegradable materials. One such materials is made
from the lactic acid produce during the bacterial fermentation of discarded corn stalks.
When individual lactic acid molecules are joined chemically, they form a material that
has the properties of plastics but is biodegradable, widespread production of plastic from
this material

41