Molecular Biology

and
Applied Genetics

By:Negeri Debela
(Medical Microbiologist)
Contents

Part-I

Genetics

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1. Introduction

• Historical overview of genetics

• Nucleic acids overview

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2. Transmission genetics

• Mendel’s laws of inheritance

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3. Chromosomes and heredity

• Sexual reproduction
• Mitosis
• Meiosis

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4. Chromosome morphology

• Chromosomal changes (Cytogenecis)

• Karyotyping

• Changes in chromosomal structure

• Sex determination

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5. Microbial genetics

• Microbial chromosome
• Microbial DNA
• Replication
• Transcription
• Translation

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 Part-II

Molecular biology

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1. Introduction

• Define terms in molecular biology

• Historical overview of molecular biology
• Overview of cellular structure and function Biologically
important molecule
• Cellular genetic components
• The central dogma of molecular biology

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2. DNA as Primary Genetic Material

• DNA structure and function

• DNA Replication and enzymes involved

• DNA Damage and Repair

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3. RNA and primary Gene Expression

• Types of RNA molecules & synthesis (Initiation, Elongation &

Termination mechanism)

• Prokaryotic transcription

• Eukaryotic transcription and transcription factors

• Chromatin structure and eukaryotic transcription

• Post-transcriptional events; RNA splicing; capping; &polyadenylation

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4. Genetic code and its translation

• Translation: Prokaryotic Vs Eukaryotic

• Genetic code

• Protein Structure related to function

• Post-translation Modification

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5. Regulation of gene expression

• Regulation of Translation

• Strategies for controlling gene-expression

• Regulation of gene expression in Prokaryotes

• Regulation of gene expression in eukaryotes

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6. Mutation

• Types of mutation

• Mutagenic agents

• How mutation affects the genetic code?

• Repair of Mutation

• Mutant isolation and detection

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7. Gene Transfer and Transposable Genetic Elements

• Extra chromosomal elements

• Plasmid
• Transposons

• Method of study Gene Transfer

• Conjugation
• Transformation
• Transduction

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 8. Molecular Techniques

• Recombinant DNA technology/genetic engineering

• Cloning and Cloning vectors

• Steps in cloning

• Gene cloning tools (Plasmids, restriction enzymes, etc)

• Gene Libraries

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8. Molecular Techniques cotn‘d.....

• Extraction of DNA, RNA from cells,

• Gel-electrophoresis
• Southern, Northern, Western Blot
• In Situ Hybridization
• PCR

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8. Molecular Techniques cotn‘d.....

• DNA Hybridization
• Sequencing
• RFLP and SNP
• Molecular markers

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Reference Books

1. Robert F. weaver, Philip W. Hedrick.Genetics.

2. Benjamin Lewin: Genes VI andabove
3. P.K. Gupta: Cell and MolecularBiology
4. Albert‗s Molecular Biology of thecell.

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Assessment methods

 Final Written Examination: 50%

 Test 1: 20%
 Test 2: 15%
 Assignment: 10%
 Lab reports: 5%

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Introduction

Genetics

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Genetics

• Genetics-from ancient Greek, genetikos, “genitive” and that from

genesis, “origin”

• Genetics defined as the study of heredity and variation in living

organism

• Genetics study include from the distribution of human traits in a

family to the biochemistry of the genetic material, like DNA, and RNA

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Heredity
• Heredity describes how some traits are passed from parents to their
children.

• Traits can be physical, such as eye colour, blood type or a disease, or

behavioural.

• The traits are expressed by genes, which are small sections of DNA that
are coded for specific traits.

• Genes are found on chromosomes.

• Humans have two sets of 23 chromosomes—one set from each parent.

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 Classification of Genetics

I-Classical Genetics II-Molecular Genetics

• Mendelian genetics • Based on classical approaches
• Aka transmission genetics • The study of the structure and

• Study of patterns of inheritance of function of genetic materials

genetic traits from one generation • Most modern genetic studies are
to the next. molecular

• Breeding experiments

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Classification of Genetics cotn’d….

III-Evolutionary genetics

• Focused on mechanism of evolutionary changes

• Change of gene frequencies in population over time

• Charles Darwin, famous in the area of evolutionary genetics

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History of Genetics

• Why Do Related
Individuals Resemble
Each Other?

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History of Genetics cotn’d….

• In the 19th century, it was commonly believed that an organism’s

traits were passed on to offspring in a blend of characteristics
‘donated’ by each parent.

• Heredity was poorly understood in general, and the concept of a

gene did not exist at all.
• The identity of the hereditary substance was unknown until 1940

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"Blending" inheritance

• (Aristotle)
• Offspring traits were a 50/50
mixture of the traits of each
parent.

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 Inheritance of Acquired Traits

• Lamarck (1744 –1829)

• Proposed that characters acquired
by individuals of one generation
are transmitted to those in the
next generation.
• Use or disuse of a structure would
cause the structure to become
Giraffe’s neck gets longer during her life because she keeps
more or less developed, stretching it to reach for leaves. Now (by the Lamarck theory)
she will have children with longer necks!
respectively. Lamarckism

01.12.2021 A weightlifter acquiresMol.

larger
Biologymuscles through constant exercise.
lecture, By Negeri 29
Particulate inheritance
Gregor Mendel (1822-1884)
• Gregor Mendel developed the
science of genetics.
• Mendel performed his experiments
in 1857.
• Principles established by Mendel
form the basis for genetics, the
science is often referred to
as Mendelian genetics.

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 Mendelian genetics

• The hybrids will have blending of

the parental traits
• Hybridization experiments with
pea plants
• Mendel cultivated and tested
some 5,000 pea plants.
• Systematically recorded results
of crosses

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 An Experiment with Tall and Short Pea Plants

• When tall plant crossed with short plant,

he always got a tall plant

• This confirmed earlier observations that

both parents contribute equally

• He then allowed hybrids to self pollinate.

• He ended up with ¼ Short plants and

¾ Tall plants

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An Experiment with Tall and Short Pea Plants

• He observed that the hybrids showed only one of the two

parental traits.

• Mendel’s results clearly disproved the blending mechanism of

inheritance

• Suggested a particulate theory of inheritance.

Discrete "particles" are passed from parents to offspring=>gene!

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Mendelian genetics

• Mendel believed that factors pass from parents to their

offspring, but he did not know of the existence of DNA.

• Modern scientists accept that genes are composed of segments

of DNA molecules that control discrete hereditary
characteristics.

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 DNA

• 1953 - James Watson & Francis Crick

• Understanding of mutation

• Relationship between DNA and proteins

• 1959 – “Central Dogma”

• DNA  RNA  protein

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 Chromosome
Condensed
chromosome
• dsDNA molecule packaged by
histone & scaffold proteins

30nm fiber

DNA double helix Nucleosome

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 How is DNA packed into chromosomes?

• DNA + histone core = nucleosome

scaffold supercoil => chromatin thread => chromosome

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 Chromatin

• The genome consists of multiple

strands of DNA packaged as
chromatin.
• Nucleosomes=>The DNA
strand is wound on histone
octamers, each composed of
four histone dimers.
• This DNA/histone complex is
known as a nucleosome.

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Chromatin

• DNA bound to both histone

• Chromosome
• 1/3 =>DNA

• 2/3=>Protein

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Gene
• Hereditary determining factor
• Continuous segment of DNA,
sequence of DNA
Maternal Chromosome # 7

• Locus – the position on a

chromosome of a particular Paternal Chromosome # 7

DNA sequence (gene)

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Alleles

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Dominant and recessive allele

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Genome

• Collection of all genetic

material of organism
• Each human cell contains
approximately 2 meters of DNA
• 3.2 × 109 nucleotide pairs
• Number of genes coding for
proteins=> Approximately
21,000

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Genome

• The human genome would

extend 3200 km
(approximately 2000 miles)

• Far enough to stretch across

the center of Africa, the site
of our human origins (red line
in B).

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Genotype & Phenotype

• Genotype
• Set of alleles present in the
genome of an organism

• Phenotype
• Result of Gene Expression
• Genes (DNA) are transcribed into
mRNA=>mRNA is translated into
protein

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The science of genetics founded by Mendel

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 The Relevance of Genetics

• Genes affect every aspect of our lives, both internally and

externally.

• It is difficult to imagine any aspect of life that does not have

something to do with genes.

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 Genetics and Agriculture

• The fastest way to increase

expression of a desirable,
recessive trait is via
inbreeding, mating close
relatives who share that trait.

• Extensive inbreeding can produce

livestock or crops that are
genetically uniform.

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 Genetics and Commerce

• Many Genetically Modified

Organisms (GMOs) can improve
human quality of life.
•Microbes can be genetically altered
to
•produce insulin
•produce hormones
•produce chemicals used to
produce medicines

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 Genetics in Society

• Powerful and controversial, genetic

testing is the process of sequencing
short DNA segments that are unique
to individuals or groups of related
individuals.

• These "DNA fingerprints" can be

used to
• Identify persons of interest from a crime
scene.
• Determine paternity of a child ……

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 Genetics in Medicine

• Faulty genes responsible for

heritable disorders
• Gene therapy is the
replacement of a malfunctioning,
mutant gene with a normal,
functional version.
• The CRISPR-Cas9 System
holds promise of a new era of
gene therapy.

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 Nucleic acids overview

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Nucleic acids overview

• Nucleic acids are molecules that store information for cellular

growth and reproduction

• There are two types of nucleic acids:

- deoxyribonucleic acid (DNA) and ribonucleic acid

(RNA)

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 Nucleic acids in human cells

• 6 billion bases in the human genome

• 2-5 % code for genes
• 35.000 to 70 000 genes in human DNA.
• The vast majority of human DNA is found in the cell
nucleus

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Chromosomal DNA: Carries the blueprint for life

• Look at each other!

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Chromosomal DNA: Carries the blueprint for life

• 99.9% identical to other humans, 98% to chimp!

• Each cell contains roughly 2 meters of DNA.

• Humans have roughly 100tril cells

• very tightly packed

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 Deoxyribonucleic acid,
Nucleic acid DNA

Ribonucleic acid, RNA

• DNA and RNA are nucleic acids, long, thread-like polymers made up of a
linear array of monomers called nucleotides

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Nucleotides

• A nucleotide consists of a
nitrogenous base, a pentose
4
5N
sugar and a phosphate group. 3

2 6
O 1
N
5’
O P O C O
O 4’ 1’

3’ 2’

OH
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 DNA vs RNA

• Ribonucleotides have a 2’-OH

• Deoxyribonucleotides have a 2’-H

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 Pentose Sugars

• There are two related pentose

sugars:
- RNA contains ribose
- DNA contains deoxyribose

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 Bases

• Purines :
– Adenine (A)
– Guanine (G)
• Pyrimidines :
– Cytosine (C)
– Uracil (U)
– Thymine (T)

DNA: A,G,C,T
RNA: A,G,C,U
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 Nucleosides =ribose/deoxyribose + bases
•The bases are covalently attached to the 1’ position of a pentose sugar ring,
to form a nucleoside

Glycosidic bond

R
Ribose or 2’-deoxyribose

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Nucleotides = nucleoside + phosphate
Phosphate ester bonds

• A nucleotide is a
nucleoside with phosphate
groups bound covalently to
the 5’ -position.

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 DNA double helix
Essential for replicating DNA and
•Two separate strands transcribing RNA
•Antiparellel (5’3’
direction) 3’
5’
•Base pairing: hydrogen
bonding that holds two
strands together

•Complementary (sequence)

• Sugar-phosphate backbones:
outside

• Base pairs : inside

•Watson and Crick , 1953

3’
5’
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 DNA double helix

4
32 1
7 6
8 5 1
9
4
3 2

A:T G:C

Base pairing

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 How does the DNA acquire stable
configuration?

 Stable configuration can be maintained by

hydrogen bond and base stacking force
(hydrophobic interaction).

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 Structures and functions of RNA

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 What is RNA?

• Ribonucleic acid
• Ribonucleotides (Ribose, base, & phosphate)

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RNA structure

• RNA molecules are largely

single-stranded but there are
double-stranded regions.

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 How many Types of RNA do you know?

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Types of RNAs

• Coding: messenger RNA (mRNA)

• Non-coding:
• Ribosomal RNA (rRNA)
• Transfer RNA (tRNA)

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Messenger RNA( mRNA)

• Function: the carrier of

genetic information from
DNA for the synthesis of
protein.

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 Transfer RNA (tRNA)

• They make up 15% of the

RNA in the cell.

• Function: Transport
amino acids to ribosomes
for assembly into proteins.

• There are at least 20

types of tRNA in one cell.
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 Ribosomal RNA (rRNA)

• Components of ribosomes.

• They make up 80% of the RNA in the cell.

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 Ribosomes

• Ribosomes are cytoplasmic structures that synthesize protein,

composed of RNA (2/3) and protein (1/3).

• The ribosomes of prokaryotes and eukaryotes are similar in

shape and function.

• The difference between them is the size and chemical

composition.

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 Summary
Base（purine, pyrimdine）+ribose（deoxyribose)
N-glycosyl linkage

Nucleoside+phosphate
Phosphoester linkage

Nucleotide
Phosphodiester linkage
Nucleic acid
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