Lesson 1: Pedigree Analysis

Relevant vocabulary

Pedigree. Making use of diagrams showing the ancestral relationships and transmission of

genetic traits over several generations in a family

Proband. The individual in the pedigree that led to the construction of the pedigree. For

example, a couple consults a medical geneticist because they have an offspring who is

afflicted with a disease and they want to find out the mode of transmission of this disease.

When the medical geneticist constructs the pedigree, the offspring will be labeled as the

proband. Through the pedigree, the probability of having other affected children may be

determined.

Law of Segregation (1st Mendelian Law). For every trait governed by a pair of alleles,

these alleles segregate or separate during gamete formation in meiosis

Law of Independent Assortment (2nd Mendelian Law). A pair of alleles for one trait will

segregate or separate independently of another pair of alleles for another trait during

meiosis

Autosomal trait. A trait whose alleles that control it are found in the autosomes (body

chromosomes/ non-sex chromosomes)

Genotype. The gene pair an individual carries for a particular trait symbolized with a pair

of letters. By convention, uppercase letter (eg. A) for a dominant allele and lowercase

letter (eg. a) for the recessive allele. Any letter in the alphabet may be used

A. For a diploid organism with two alleles in a given gene pair, genotypes may be

written as:

i. Homozygous dominant, i.e. with two dominant alleles (DD)

ii. Heterozygous, i.e. with a dominant and recessive allele (Dd). The individual will

show the dominant phenotype.

iii. Homozygous recessive, i.e. with two recessive alleles (dd)

Phenotype

A. The observable trait of an individual based on its genotype. Examples: red flower, curly

hair, blood types ( i.e. the blood type is the phenotype)

B. For a typical Mendelian trait, phenotypes may either be:

i. Dominant. A trait that requires at least one dominant allele for the trait to be

expressed, e.g. Dd

ii. Recessive. A trait that requires two recessive alleles for the trait to be expressed

Phenocopy. A trait that is expressed due to specific environmental conditions (i.e. having

hair that is dyed of a different color) and is not due to the genotype.

Identical twins. Also known as monozygotic twins, which are derived from a single

fertilization event. After the first cleavage or cell division of the zygote, the cells or

blastomeres separate and become independent blastocysts implanted in the mother’s

uterus.

Fraternal twins. Twins that are derived from separate fertilization events (two eggs

fertilized by two sperms) within the fallopian tube, resulting in two separate zygotes; also

known as dizygotic twins

Symbols for creating pedigrees

I. Male (square) vs female (circle)

II. Affected (shaded) vs unaffected (unshaded) individual

III. Marriage/mating line (line connecting mates) vs. sibship line

(line connecting siblings)

IV. Fraternal twins (one birthline branching out into the

individual twin) vs. identical twins (same as fraternal twins but

with a horizontal bar connecting the branches)

V. Generation (Roman numerals) vs. individuals in the same

generation, counting left to right (designated by Hindu-

Arabic numerals)

VI. Proband (arrow)

What to expect in a human pedigree

For autosomal dominant trait: Two affected individuals can

have a normal offspring

For autosomal recessive trait: Two affected individuals can

NEVER have a normal offspring

Example:

I. Identify the mode of inheritance.

II. Write down the genotypes of specific individual
III. Compute for the probability of having an affected offspring
A. Look at the family of IV-9 and IV-10. If the trait is dominant, is it possible for them to have an affected offspring?
(Answer: NO. If the trait is dominant, then unaffected
individuals are homozygous recessive. Two recessive
individuals CANNOT produce a dominant offspring.)
B. If the trait is recessive, is it possible for IV-9 and IV-10 to have an unaffected offspring?
(Answer: YES. This can happen if both parents are
heterozygous for the trait, which means they can each
give a recessive allele to produce a homozygous
recessive offspring.)
C. Based on your answers in A and B, is the trait dominant or recessive?
(Answer: RECESSIVE)
D. Give the genotypes of the following
i. IV-9 (Answer: Dd)
ii. IV-10 (Answer: Dd)
iii. V-1 (Answer: DD or Dd)
iv. I-1 (Answer: dd)
v. I-2 (Answer: Dd)
E. If IV-9 and IV-10 were to have another child, what is the probability that they will have an affected offspring?
(Answer: 1/4 or 25% following the Mendelian ratio from a
hybrid cross)
Lesson 2: Sex Linkage and
Recombination
I. Sex linked trait. The gene (pair) that determines a character (e.g. hemophilia) is located

on the sex chromosomes

II. X-linked trait. A sex-linked trait is where the gene or allele for the trait is found on the X

chromosome

III. Color blindness. An X-linked recessive trait where a affected individual could not

distinguish red from green color (red green color blindness)

IV. Hemophilia. An X-linked recessive trait where an affected individual suffers from delayed

blood clotting during injuries because of the absence of certain blood clotting factors

V. Y-linked trait. A sex-linked trait where the gene or allele for the trait is found on the Y

chromosome

VI. Hypertrichosis pinnae auris. A Y-linked trait where affected males have hair growing from

their external ears

VII. Other sex-related traits.

A. Sex-influenced trait- Any trait in a diploid organism whose expression is affected by

an individual’s biological sex; a trait that occurs at a higher frequency in one sex over

the other

B. Sex-limited trait- Any trait in a diploid organism whose expression is limited to just

one biological sex

C.
Sex-linked traits

• In humans, males and females are represented by different

sex chromosomes

• Females have two X chromosomes in the nucleus of their

cells.

• Males have one X chromosome and one Y chromosome in

the nucleus of their cells.

• Depending on whether the trait is dominant or recessive, the

expression pattern of the trait differs in males and females

• Colorblindness in humans as an example of sex-linked trait

• The alleles responsible for colorblindness is found on the X

chromosome only

• The dominant allele is the normal allele; the recessive allele

causes colorblindness

• Females need two copies of the recessive allele, one from

each of the two X chromosomes, for the trait to be

manifested. If they only have one copy of the recessive

allele, they have normal color vision. However, they are

carriers for the trait in that they may pass it on to their

offspring.

• Males only need one recessive allele in their sole X

chromosome for the trait to be expressed.

• There is difference in expression between males

and females for Y-linked traits. (Since the allele is found only

in the Y chromosome, and since only males have Ychromosomes,

then only males will express the trait.

Females CANNOT express Y-linked traits.)

• Hypertrichosis pinnae auris as an example of a Y-linked trait

• If a male has the allele responsible for the trait, then his Y

chromosome will possess that allele. Since he will pass on his

Y chromosome to his sons, then all his sons will inherit the

trait, and they, in turn, can pass on the allele to their sons.

Sex-influenced trait

• Hormonal or physiological differences between the sexes

cause differences of expression of certain genes

• Baldness in humans as an example of a sex-influenced

trait. See Table 1 how baldness is hypothesized to be

expressed by a single pair of alleles, with B as the

dominant allele for baldness and b as the recessive

normal allele.

Sex-limited traits

• Hormonal or physiological differences between sexes

may limit the expression of some genes to one biological

sex only

• Functional mammary glands as an example of a sexlimited

trait. Only females can express functional

mammary glands that produce milk immediately after

giving birth.

• Note that baldness behaves like a dominant trait in

males in that only one dominant allele is needed for

baldness to be expressed. On the other hand, the trait

behaves like a recessive trait in women in that they need

both dominant alleles to be present for baldness to be

expressed.
Lesson 3: Modification to Mendel’s
Classic
Ratios
I. Co-dominance - When two contrasting alleles are present in

the same locus or trait (heterozygote genotype), then the

phenotype expressed is a “blend” of the two extreme

phenotypes. The two genes interact and the offspring shows

the effects of both alleles.

II. Incomplete dominance - When two contrasting alleles are

present in the same locus or trait (heterozygote genotype),

then both alleles are expressed in the same phenotype

III. Multiple alleles - When there are more than two types of

alleles for a given locus or trait, this will result in more than

two kinds of phenotypes that may be expressed for that

trait.

Incomplete dominance. The heterozygote genotype is expressed as a

distinct phenotype (a “blend” of the two extreme phenotypes). In this case, the phenotypic

ratio is the same as the genotypic ratio

I. Use snapdragon plants (Antirrhinum majus) as example (see figure 1).

A. RR – red flowers

B. Rr – pink flowers

C. rr – white flowers

Co-Dominance. The heterozygote genotype is expressed as a distinct

phenotype (both extreme phenotypes are expressed at the same time). Similar to incomplete

dominance, the phenotypic ratio is the same as the genotypic ratio.

I. Use human MN blood typing as an example

A. MM – type M
B. MN – type MN

C. NN – type N

Multiple alleles. There are more than two types of alleles, and the

relationship of each allele with respect to others will determine the number of phenotypes

that may be expressed.

I. Use coat color in rabbits as example (see figure 2)

A. There are four different types of alleles in rabbits: C (Agouti), Cch (Chinchilla), Ch

(Himalayan), and c (Albino), with the following dominance hierarchy: C> Cch>Ch> c.

B. The following genotypes will have the corresponding phenotypes in coat color:

i. CC – Agouti

ii. CCch – Agouti

iii. CCh – Agouti

iv. Cc – Agouti

v. CchCch – Chinchilla

vi. CchCh – Chinchilla

vii. Cchc – Chinchilla

viii.ChCh – Himalayan

ix. Chc – Himalayan

x. Cc – Albino

II.Use ABO blood typing in humans as example

i. There are three different types of alleles A (or IA), B (or IB) and O (or i)

ii. The following genotypes will have the following blood types (phenotypes):

iii. AA (or IAIA) – Type A

iv. AO (or IAi) – Type A

v. BB (or IBIB) – Type B

vi. BO (or IBi) – Type B

vii. AB (IAIB) – Type AB

viii.OO (ii) – Type O

Incomplete dominance in snapdragons, Antirrhinum Coat color in rabbits. The trait is controlled b
majus. The multiple alleles with

cross involving homozygote red flowers (RR) and the following dominance hierarchy: C (Agouti) >
homozygote white Cch (Chinchilla) >

flowers (rr) will yield a heterozygote (Rr) that Ch (Himalayan) > c (Albino).
expresses a different

phenotype, which is pink flowers. The cross between

pink-flowered

individuals will produce offsprings where the

genotypic ratio also

becomes the phenotypic ratio (25% red: 50% pink:

25% white).
Lesson 4: Molecular Structure of DNA,
RNA, and Proteins
1. The building blocks of any nucleic acid are the nucleotides.

2. A nucleotide is composed of a phosphate group (with negative charges), a sugar portion and

an N-base.

3. The sugar in DNA is deoxyribose while the sugar in RNA is ribose. Explain the difference

through a visual aid.

4. DNA and RNA are polynucleotides. N-bases are either purines or pyrimidines. Purine bases

are Adenine (A) and Guanine (G). Pyrimidines are Cytosine (C), Thymine (T, in DNA only) and

Uracil (U, found only in RNA)

5. Specific base pairings occur in DNA. A pairs with T; G pairs with C

6. DNA is double stranded while RNA is single stranded with Uracil instead of Thymine.

7. Main Functions:

I. DNA: repository of genetic information; sequence of bases encodes the blueprint for life

processes

II. RNA: information in the form of base sequence is transformed (transcribed) into mRNA,

tRNA and rRNA. DNA is the template copied into RNA by base pairing. G with C; A with

U.

III. Protein: functional products of genes; executes cellular functions

8. The four structural levels of proteins are: 1.Primary- sequence of amino acids in the

polypeptide chain; 2. Secondary- when the polypeptide chains form a helix or a pleated sheet

structure; 3. Tertiary- coiling of the polypeptide, combining helices and sheet forms; 4.

Quaternary- the association of two or more polypeptides in space

Lesson 5: DNA Replication and Protein
Synthesis
1. DNA replication or DNA synthesis. DNA strands separate and serve as templates for the

production of new DNA molecules.

A. The following are features of replication:

i. Semiconservative- the resulting DNA consists of one old and one new strand

ii. Base pairing is maintained; Adenine pairs with Thymine, Guanine pairs with Cytosine

iii. New DNA molecules are produced in the 5’ to 3’ direction

iv. Semidiscontinuous. The leading strand is synthesized in a continuous manner (5’ to 3’)

while the lagging strand is produced discontinuously in short stretches called Okazaki

fragments.

B. In lagging strand synthesis, there is a need for a primer terminus which is provided by an

RNA molecule. RNA is synthesized by a primase or RNA polymerase. The 3’OH of the

RNA is where new DNA nucleotides are added thus new DNA is built in the 5’ to 3’

direction.

C. Enzymes in replication are as follows: 1. helicase; 2. gyrase; 3. SSB (single strand binding

proteins); 4. primase or RNA polymerase; 4. DNA polymerase and 5. DNA ligase.

2. Transcription or RNA synthesis. DNA is unwound and one strand is used as template for the

production of an RNA molecule. An RNA polymerase makes RNA in the 5’ to 3’ direction.

Specific regions in the DNA called promoters allow the binding of transcription factors which

make possible the binding of RNA polymerase. Three major types of RNA are: messenger

RNA (mRNA); transfer RNA (tRNA) and ribosomal RNA (rRNA).

3. Translation or protein synthesis. This occurs in the ribosome. Basic ingredients are the

various types of RNAs produced in transcription and some proteins or enzymes. The mRNA

contains triplets of bases called codons that specify an amino acid, eg. UUU-phe. Various

tRNAs carry amino acids from the cytoplasm to the actual site of translation in the ribosome. A

tRNA has an anticodon that pair with a codon in the mRNA. Different rRNAs combine with

ribosomal proteins to make up the subunits of a ribosome. A functional ribosome has a small

and a large subunit.

In bacteria, transcription and translation may be simultaneous. In eukaryotic cells, mRNA,

tRNA and rRNA travel from the nucleus to the cytoplasm through the nuclear pores. RNAs

may undergo processing. Some unnecessary parts like introns are removed. In eukaryotic

mRNA, a 5’ cap and a 3’ poly A tail are added. Coding regions of mRNA are called exons.

They specify functional protein products.

Lesson 6: Genetic Engineering
Genetic Engineering

1. Classical breeding practices focus on the mating of organisms with desirable qualities.

2. Genetic engineering involves the use of molecular techniques to modify the traits of a target

organism. The modification of traits may involve:

I. introduction of new traits into an organism

II. enhancement of a present trait by increasing the expression of the desired gene

III. enhancement of a present trait by disrupting the inhibition of the desired genes’

expression.

3. A general outline of recombinant DNA may be given as follows:

I. cutting or cleavage of DNA by restriction enzymes (REs)

II. selection of an appropriate vector or vehicle which would propagate the recombinant

DNA ( eg. circular plasmid in bacteria with a foreign gene of interest)

III. ligation (join together) of the gene of interest (eg. from animal) with the vector ( cut

bacterial plasmid)

IV. transfer of the recombinant plasmid into a host cell (that would carry out replication to

make huge copies of the recombined plasmid)

V. selection process to screen which cells actually contain the gene of interest

VI. sequencing of the gene to find out the primary structure of the protein
4. After outlining the key steps in recombinant DNA, the teacher can proceed to describe the

ways in which these plasmids may be introduced into host organisms.

Biolistics. In this technique, a “gene gun” is used to fire DNA-coated pellets on plant tissues.

Cells that survive the bombardment, and are able to take up the expression plasmid coated

pellets and acquire the ability to express the designed protein.

Plasmid insertion by Heat Shock Treatment. Heat Shock Treatment is a process used to

transfer plasmid DNA into bacteria. The target cells are pre-treated before the procedure to

increase the pore sizes of their plasma membranes. This pretreatment (usually with CaCl2) is

said to make the cells “competent” for accepting the plasmid DNA. After the cells are made

competent, they are incubated with the desired plasmid at about 4°C for about 30min. The

plasmids concentrate near the cells during this time. Afterwards, a “Heat Shock” is done on

the plasmid-cell solution by incubating it at 42°C for 1 minute then back to 4°C for 2 minutes.

The rapid rise and drop of temperature is believed to increase and decrease the pore sizes in

the membrane. The plasmid DNA near the membrane surface are taken into the cells by this

process. The cells that took up the plasmids acquire new traits and are said to be

“transformed”.

Electroporation. This technique follows a similar methodology as Heat Shock Treatment, but,

the expansion of the membrane pores is done through an electric “shock”. This method is

commonly used for insertion of genes into mammalian cells.