LECTURE 5:

DNA, RNA & PROTEINS

The molecules of life

Today…
• DNA structure and replication

• RNA
– Transcription
– Translation

• Protein synthesis
– Amino acids
 DNA
• Deoxyribonucleic Acid (DNA)
• is the blueprint for life:
• contains __________________________
 DNA structure
• DNA is a polymer of
nucleotides
Each nucleotide composed of
Sugar___________
Phosphate_______
Nitrogen Base____
 Four DNA bases
• Four kinds of nitrogenous bases:
• Purine bases

• Pyrimidine bases
DNA: Complimentary base pairing
• Adenine pairs with ____

A T
• Cytosine pairs with _____

C G
 DNA STRUCTURE
• DNA is a _Double_ helix
• Discovered by
Watson and Crick, 1953
 DNA REPLICATION
(in the nucleus)
• Each DNA strand
becomes a Template,
parent strand becomes apart
• Proper base-pairs are
assembled on that template
 DNA replication
• Nucleotides are connected together to make
a new strand that is complimentary_ to the
old strand.
• The new double strand is
identical_ to the old double strand
• This is called ________________________
replication
 A-T vs. G-C bond
• A-T is a double bond…

• C-G is a –triple -- bond (stronger)…

• DNA double strand can separate into 2 single

strands when heated.
• Which strand would require more heat (more
energy) to separate: an A-T rich or a C-G rich
double strand?
 RNA structure and synthesis
• RNA: Single stranded___________________
• Is very similar to DNA
(repeating subunits, nucleotides).

• Difference between RNA and DNA:

Each nucleotide contains a different sugar:
_ribose___________ instead of deoxyribose.
Bases are A, G, C, and U (Uracil_)
A pairs with U______; G pairs with _C_____
 RNA
• RNA is _Single________ stranded
and shorter

• RNA is less stable than DNA:

RNA doesn’t persist in the cell for long
(sometimes it exists for a few seconds),
whereas DNA can persist for the life of the
cell.
 CENTRAL DOGMA
transcription

DNA translation

RNA
Proteins

• 3 different RNA molecules involved in

protein synthesis: mRNA, rRNA, tRNA__
• TRANSCRIPTION: RNA synthesis from
DNA.
For Teaching & academic instruction only
 For Teaching & academic instruction only

Fig. 2. The arrows show the situation as it seemed in

1568. Solid arrows represent probable transfers, dotted Fig. 3. A tentative classification for the present day. Solid arrows
arrows possible transfers. The absent arrows (compare show general transfers; dotted arrows show special transfers.
Fig. 1) represent impossible transfer postulated by the Again, the absent arrows are the undesired transfers specified by
central dogma. They are three possible arrows starting central dogma.
from the protein
For Teaching & academic instruction only
For Teaching & academic instruction only
 Transcription: DNA → RNA
• Transcription occurs in the _Nucleus________
• mRNA carries the message about what type of
protein to make from the DNA in the nucleus to
the ribosome
• The nucleotide sequences of RNA and DNA are
the same (except in RNA __Uracil_____ is used
instead of thymine)
• mRNA is synthesized from DNA using base
pairing
• DNA unwinds in a section
• RNA _polymerase__ attaches at the promoter sequence
of DNA, and it moves along the DNA, unzipping the
strands – this allows for one mRNA molecule to be
formed.
• During _Transcription__, a molecule of messenger RNA
is formed as a complementary copy of a region on one
strand of the DNA molecule
 Transcription
• Once mRNA is formed, enzymes in the nucleus
remove the
• Introns _(Non-coding part)_ and leave the
• Exons _(Coding part)__
 The Genetic Code
• Each 3 consecutive bases on the mRNA is a code word,
codon, that specifies an amino acid.
• The genetic code consists of _Triplet____codons,
• but only__code single amino acids.
• Three codons act as
signal terminators
(UAG_,UAA, UGA)

• One codon, AUG, codes

for methionine, and is also
the _start_____ signal for translation.
 20 Amino Acids
There are _20___ amino
acids – they are like the
‘bricks’, or building
blocks to make
all _Proteins_______
 Translation: RNA → Protein
• Translation: synthesizing a _Protein___ from
amino acids, according to the sequences of the
nucleotides in mRNA.
• Occurs at the Ribosomes _, in cytoplasm of cell
• Ribosomal RNA, rRNA, is needed for protein
synthesis – helps mRNA bind to the
_Ribosome_
• _tRNA__________ brings specific amino acids
to the ribosome to be assembled as proteins.
• _Ribosomal____ RNA,
rRNA, joins with a
Translation
number of proteins to
form ribosomes
• Ribosomes are the
sites of _Protein
Synthesis_
• Ribosomes consist of
a large subunit and
a small subunit.
mRNA binds to the
small___ subunit.
 Transfer RNA (tRNA)
• Transport molecule
that carries specific
amino acids ____ to a
ribosome
(80 nucleotides long)
• Folded

• Each tRNA recognizes

the correct codon on
the mRNA molecule
Translation
 Steps in Translation
1. mRNA leaves the nucleus and migrates to ribosome
2. mRNA binds to small ribosomal subunit
3. tRNA brings an amino acid to the ribosome, where
anticodon on the tRNA binds to the codon of the
mRNA
4. The amino acid bonds to its adjoining amino acid to
form a growing polypeptide molecule
5. The tRNA without the amino acid is released from the
ribosome
6. Other tRNA’s bring amino acids to the ribosome to
complete the protein molecule
Protein translation
 Protein synthesis

• Occurs in the cytoplasm of the cell

 Protein synthesis
• Amino acids are the repeating sub-units of
protein molecules.
• Amino acid order determines the protein
• 20__ amino acids exist in all life forms
• Order _of amino acids is important, determines
the 3-dimensional shape of the molecule.
• Structure of the protein determines its Activity_
 Proteins
• Biological activity (function) of proteins
depends largely on its 3-D structure
Summary:
 Genomic
Geography

• In Cell Nucleus: RNA is produced by transcription.

• RNA is single-stranded; substitutes the sugar ribose for deoxyribose and the base uracil for thymine
• Messenger RNA or mRNA, conveys the DNA recipe for protein synthesis to the cell cytoplasm.
• mRNA binds to ribosome, each three-base codon of the mRNA links to a specific form of
transfer RNA (tRNA) containing the complementary three-base sequence.
• This tRNA, in turn, transfers a single amino acid to a growing protein chain.
• Each codon directs the addition of one amino acid to the protein. Note: the same amino acid can be
added by different codons; in this illustration, the mRNA sequences GCA and GCC are both
specifying the addition of the amino acid alanine (Ala).
 Important
• Both DNA and RNA have a direction: one end is
the 3’ the other is the 5’ end.
• Thus, codons are read in one direction only.

• Also, note there is redundancy in the genetic code:

the different sequences can specify for the same
amino acid.
Example: Leucine is coded by six codons: UUA,
UUG, CUU, CUC, CUA, and CUG
 When things go wrong…
• Mutations: changes in the DNA sequence, that may be
passed along to future generations.
• Point mutations: a single base substitution
THE CAT SAW THE RAT
THE CAT SAW THE HAT
• Deletion: a small DNA segment is lost

• Insertion: a segment of DNA is added

• Frame-shift mutation: modification of the reading

frame after a deletion or insertion, resulting in all
codons downstreams being different.
 Somatic Mutations
• Somatic______ mutations: occur in body
cells, or cells that do not lead to gametes.
• Somatic mutations that occur in leaves, roots
or stems are usually not passed on to future
generations…
• UNLESS the plant reproduces ____________
Gregor George Mendal (1822-1884)

1. Austrian Monk, mathematician turned biologist and

become “Father of Genetics”

2. Develop a simple set of rules to accurately predict

patterns of heredity which forms the basics of genetics

3. Years later we found that traits are determined by

genes encoded in DNA
Heredity
1. Transmission of traits from parents to offspring, before DNA was
discovered it was one of the great mysteries of science!

2. Modeled experiments after British farmer T.A. Knight who bred garden
peas and concluded purple flowers show a stronger tendency to appear
than white flowers

1. Mendel used a mathematical approach and counted the number of each

kind of offspring
Why did Mendel choose peas?
1. Distinct, Observable Traits: Pea plants exhibit clear, easily distinguishable traits, such as
flower color (purple or white), seed shape (round or wrinkled), and seed color (yellow or
green). This made it straightforward for Mendel to track and analyze trait inheritance.
2. Controlled Mating:
Peas are self-pollinating, meaning they normally fertilize themselves.
However, Mendel could easily control cross-pollination by manually transferring pollen
between plants, allowing him to design specific crosses and isolate variables. (Emasculation)

Self-pollination Cross-
pollination
1. Short Life Cycle: Pea plants have a relatively short generation time, meaning Mendel could observe
multiple generations in a relatively short period. This was crucial for studying patterns of inheritance over
successive generations.

2. Large Number of Offspring: Pea plants produce many seeds per plant, giving Mendel a large sample size
for statistical analysis of trait inheritance, which increased the accuracy of his conclusions.
Mendel’s Experimental Design and results

F1 The recessive
traits(Green seed)
disappears, The (Yellow
Seed) expressed trait is said
to be dominant

F2 The recessive trait

reappears!!
Mendel obtained a 3:1 ratio
of dominant to recessive for
each trait of the F2
generation!
Monohybrid
Cross
Law of
Segregation
Law of segregation: Meiosis

Cross pollination

All heterozygous plant in F1 generation

Dihybrid
Cross
 y y
Law of Independent Y Y

Assortment
R R r r

YyRr YyRr

YR YR yr yr Yr Yr yR yR
At the end of this lesson you should be able to
1. Define Genetic Engineering

2. Outline the process of genetic engineering

involving some or all of the following: isolation,
cutting, transformation, introduction of base
sequence changes and expression

3. Know three applications: one plant, one

animal, one micro-organism
The simple addition, deletion, or
manipulation of a single trait in an organism
to create a desired change.
 Manipulation and
alteration of genes
 Three applications:
one plant, one animal,
one micro-organism

 Process involving
isolation, transformation,
and expression
Is:
Artificially copying a piece of DNA from one
organism and joining this copy of DNA into the
DNA of another organism

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 It allows genes from one organism to be inserted
into a cell of a different organism of a different
species.

Examples:
– Human genes can be inserted into a
bacterium
– Human genes can be inserted into cells from
other animals
– Bacterium genes can be inserted into plant
cells
 Genetic engineering means that DNA from
different organisms can be combined

 Bacteria can be engineered to produce human

proteins

 Human genes can be inserted into other

animals
 The altered DNA is called recombinant DNA

 Recombinant DNA is joined to other

unrelated DNA in the organism

 This is called gene splicing.

- tiny segments of a gene are taken out

and replaced by different genes
1. What is Genetic Engineering?

2. What is the purpose of Genetic Reengineering?

3. Give three examples of Genetic Engineering

4. What is the difference between altered and

recombinant DNA?

5. What is meant by gene splicing?

 Organisms altered by genetic engineering.

 Genetic material changed by other than random

natural breeding

 Gene transfer
-moving a gene from one organism to another.
 'Trans-' means 'crossing from one place to
another‘

 The '-genic' bit means genes

 So it means that bits of genes from different

living things have been bolted together and
spliced into another organism to make a new
one which does something which the scientists
want it to do.
 GMO- genetically modified organism
 GEO-genetically enhanced organism

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For example
Plants that resists a particular type of weed killer
Sheep which makes some special substance in its milk.
1. What the word transgenic mean?

2. What is a transgenic organism?

3. Give examples of transgenic organisms

1. Isolation

2. Cutting

3. Ligation and Insertion

4. Transformation

5. Expression
 (a) Isolation of a specific gene from donor e.g. human

• Cells broken open

• Genetic probe added

• Reveals position of the gene of interest

Genetic probe

Position of
gene of
interest

Donor DNA
(b) Isolation of plasmid from a bacterial cell
Bacterial cell

Plasmid

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 Restriction enzymes act as molecular scissors and cut
DNA at specific sites called restriction sites

Restriction site
Restriction site
Restriction ezymes
Restriction site Restriction site
Donor DNA

Plasmid
Restriction
enzymes
© Biology Support Service 2007 18
Donor DNA

Plasmid Sticky Ends

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Ligation –rejoining cut fragments of DNA and

forming artificial recombinant molecules
 Recombinant DNA introduced into
bacterial cell

Bacterial
cell
Bacterial
chromosome

Recombinant DNA
Bacterial cell reproduces by Binary Fisson

Bacterial cell produces the polypeptide

Coded for by the donor DNA

 Donor DNA
Plasmid

1. Cut with restriction

enzymes

Donor DNA

Sticky
Ends

2. Ligase bonds
sticky ends Recombinant DNA
together
 Expression is getting the organism with the
recombinant DNA to produce the desired
protein

 When the protein is produced in large amounts

it is isolated and purified
1. What are the stages involved in GE?

2. Outline what happens in the stage isolation ?

3. What enzyme is used to cut the DNA?

4. Can you explain what happens in

transformation?

5. What is meant by gene expression?

 The human gene to clot blood has been
inserted into the DNA of sheep

 Sheep produce human clotting factor needed

for Haemophiliacs in their milk

 Goats produce a protein to treat emphysema

Production of humulin

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Used by diabetics
28
White rice and golden rice

Golden Rice – a
possible solution to
Vitamin A
deficiency.

29
 Pharming
 Gene pharming is a technology that scientists use
to alter an animal's own DNA, or to splice in new
DNA, called a transgene, from another species.
 In pharming, these genetically modified
(transgenic) animals are mostly used to make
human proteins that have medicinal value. The
protein encoded by the transgene is secreted into
the animal's milk, eggs or blood, and then
collected and purified.
 One of the first mammals engineered
successfully for the purpose of pharming was a
sheep named Tracy, born in 1990 and created
by scientists led by British developmental
biologist Ian Wilmut at Roslin Institute in
Scotland. Tracy was created from a zygote
genetically engineered through DNA injection
to produce milk containing large quantities of
the human enzyme alpha-1 antitrypsin, a
substance used to treat cystic fibrosis and
emphysema
 Xenotransplantation is the transplantation of
living cells, tissues or organs from one species
to another.
 However there are ethical issues and issues
with rejection
 There are also issues with virus transmission
from one species to another
 Porcine islet transplants are being investigated
for use in type 1 diabetes due to the shortage of
human islet cells
 It involves modifying
human DNA either to
repair it or to replace a
faulty gene.

 The idea of gene therapy

is to overcome the effects
of a mutation which
causes a genetic disease.

 Cystic fibrosis is the best

known disease where
gene therapy has been
tried.
Genetic engineering can produce very
specific and sensitive diagnostic tests for
many diseases, using engineered proteins.

This new technology is also opening up

novel ways of delivering medicines to
specific targets.
 Genetically engineered
microbes can be used to
produce the antigens
needed in a safe and
controllable way.
 The use of genetically
modified yeast cells to
produce a vaccine
against the hepatitis B
virus has been a major
success story.
 Weedkiller resistant crops
- Weeds die but the crops survive
 Vitamin A in Rice

- The gene which produces vitamin A was taken

from daffodils and put into rice to help prevent
blindness
 Bacteria can make human insulin

 This prevented many diabetics from getting an

allergic reaction to animal insulin

 Bacteria make interferon which can fight virus

infections and some cancers
1. Can you outline 3 uses of Genetic Engineering?

2. What is the purpose of Pharming?

3. Outline a use for Gene Therapy

4. What have GE modified yeast cells been used to

produce?

5. What do you feel are the ethical issues associated

with GE?
Can you…………

1. Define Genetic Engineering

2. Understand that GE alters DNA
3. Understand the function of restriction enzymes
4. Be able to explain isolation, cutting , insertion ,
transformation and expression
5. Discuss three applications of GE
6. Discuss the ethical issues of genetic
engineering
CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) is a
revolutionary genome-editing technology that allows precise modifications to
DNA in living organisms.

Originally discovered as a bacterial defense mechanism against viruses,

CRISPR works by using the Cas (CRISPR-associated) proteins, such as Cas9 or
Cas12, to target and cut specific DNA sequences guided by a programmable
RNA.

This system has transformed molecular biology, enabling gene knockouts,

corrections of genetic mutations, and advancements in medicine, agriculture,
and biotechnology.
A CRISPR region within a microbial genome
In prokaryotic CRISPR-Cas systems, CRISPR arrays consist of short, repeating DNA
sequences (repeats) interspersed with unique DNA sequences (spacers), which are
derived from fragments of invading genetic elements, serving as a record of past
infections and enabling adaptive immunity
1. The Cas1-Cas2 enzymes of the microbe
recognize and cut out a segment of foreign DNA.

2. The Cas1-Cas2 enzymes insert the DNA segment into

the CRISPR region of the bacterial genome as a spacer.
3. A spacer sequence is transcribed and then linked to a
Cas9 protein.

4. Upon reinfection by the same invader, the CRISPR-

Cas9 complex can recognize the foreign DNA sequence
and cut it to prevent complete reinfection.
 CRISPR has revolutionized biotechnology and medicine with its
diverse applications

1. Gene Knockout & Knock-in – Modifying genes for functional studies.

2. Precision Medicine – Correcting disease-causing mutations (e.g., Sickle Cell Anemia, Cystic Fibrosis).
3. Transgenic Crops & Animals – Enhancing traits like disease resistance, drought tolerance, and higher
yields.
4. CRISPR-based Gene Therapy:Treating genetic disorders like Beta-thalassemia, Leber congenital
amaurosis (LCA), Duchenne Muscular Dystrophy (DMD).
5. Oncology – CRISPR-edited immune cells (CAR-T therapy) for cancer treatment
6. Crop Improvement – Enhancing yield, resistance to pests, and stress tolerance in plants like wheat, rice,
and maize.
7. Livestock Engineering – Improving disease resistance and productivity in animals (e.g., tuberculosis-
resistant cows).