Lecture PowerPoint to accompany

Molecular Biology
Fifth Edition

Robert F. Weaver

Chapter 1
A Brief History

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
 Introduction
• 수업 개요.
분자생물학은 유전체의 복사,전사 및 번역 과정 과 같은 생명의 제현상을
분자수준에서 연구하는 학문으로서 의학, 약학, 농학, 식품공학, 환경 공학등
기초 및 응용분야에 지대한 영향을 끼쳐 관련 분야의 획기적인 진전을
유발하고 있습니다.
1953년 제임스 왓슨 과 크릭이 DNA의 이중 나선을 발견한 시점 에서 분자
생물학이 탄생 했다고 볼수 있으며, 이후 분자 생물학은 생화학과 유전학의
영역을 흡수해가며 급격하게 성장해왔습니다.
현대생물학은 기본적으로 분자 생물학을 배경으로 하고 있으며, 세포내 또는
세포간에 이루어지는 여러 가지 형태의 상호 작용들을 해석하는 과정을
기본으로 합니다. 현재는 인간 유전체 분석사업 (human genome project) 이
끝나고, 이 정보를 기반으로 해 다양한 접근 방법이 시도 되고 있습니다.

1-2
 Introduction
• Course objectives:

• 생명체 구성 과 생명 현상에 대한 분자 수준에서의 기초 지식 습득

• 원핵 및 진핵 생물의 유전자 발현 과정과 조절 기작, 단백질 번역

과정 그리고 복제 과정 등을 포함한 생명 현상 이해

• 최신 분자 생물학적 분석 방법 습득 및 이해

1-3
 Introduction
• Student responsibilities:

• Attendance: minimum effort to know your passion

• Preparation: read at least 1 time; highly recommended
• Review your own notes -text (where appropriate),
• Study (?)
• ... and ask questions

1-4
 Curriculum
• 1 week: Introduction
• 2 week: The Molecular Nature of Genes
• 3 week: An Introduction to Gene Function
• 4 week: Molecular Cloning Methods
• 5 week: Molecular Tools for Studying Genes and Gene Activity
• 6 week: The Mechanism of Transcription in Bacteria
• 7 week: Operons: Fine Control of Bacterial Transcription
• 8 week: Mid-term Exam.
• 9 week: Major Shifts in Bacterial Transcription
• 10 week: DNA;Protein Interactions in Bacteria
• 11 week: Eukaryotic RNA Polymerases and Their Promoters
• 12 week: General Transcription Factors in Eukaryotes
• 13 week: General Transcription Factors in Eukaryotes
• 14 week: Transcription Activators in Eukaryotes
• 15 week: Chromatin Structure and Its Effects on Transcription
• 16 week: Final Exam.
1-5
 Introduction
• Evaluation

• Mid-Term Exam: 45 %

• Final Exam: 45 %

• Attendance: 10 %

1-6
 A Brief History
• What is molecular biology?
– The attempt to understand biological
phenomena in molecular terms
– The study of gene structure and function
at the molecular level
• Molecular biology is a melding of
aspects of genetics and biochemistry

1-7
 1.1 Transmission Genetics
• Transmission genetics deals with the
transmission of traits from parental
organisms to their offspring
• The chemical composition of genes
was not known until 1944
– Gene - genetic units
– Phenotype - observable characteristics

1-8
Gregor Mendel: Father of
Modern Genetics

1-9
 Drawing from the Deck of
Genes
• What genetic principles account for the passing
of traits from parents to offspring?

• Blending hypothesis- genetic material from the

two parents blends together (like blue and
yellow paint blend to make green).

• Incubation hypothesis- only one parent

controlled the traits of the children.
Ex: Spermists and Ovists
1-10
 Drawing from the Deck of
Genes
• Particulate hypothesis- parents pass on
discrete heritable units (genes) that retain their
identities in the offspring

• This hypothesis can explain the reappearance

of traits after several generations

• Mendel documented a particulate mechanism

through his experiments with garden peas

1-11
 Mendel’s Experiment
• In a typical experiment, Mendel mated two contrasting,
true-breeding varieties, a process called hybridization

• The true-breeding parents are the P generation

• The hybrid offspring of the P generation are called the

F1 generation (first filial generation)

• When F1 individuals self-pollinate or cross- pollinate

with other F1 hybrids, the F2 generation is produced

1-12
 The Law of Segregation
• When Mendel crossed contrasting, true-
breeding white- and purple-flowered pea plants,
all of the F1 hybrids were purple

• When Mendel crossed the F1 hybrids, many of

the F2 plants had purple flowers, but some had
white

• Mendel discovered a ratio of about three to one,

purple to white flowers, in the F2 generation
1-13
 P Generation
(true-breeding
parents) Purple White
flowers flowers

F1 Generation
(hybrids)
All plants had purple flowers
When F1 hybrid pea
Self- or cross-pollination plants self- or cross-
pollinate, which traits
appear in the F2
F2 Generation generation?

705 purple- 224 white

flowered flowered 1-14

plants plants
 Mendel’s Laws of Inheritance
• A gene can exist in different forms called alleles
• One allele can be dominant over the other,
recessive, allele
• The first filial generation (F1) contains offspring
of the original parents
• If each parent carries two copies of a gene, the
parents are diploid for that gene

1-15
 Alleles,
alternative versions of a gene.
Allele for purple flowers

Pair of
Locus for flower-color gene homologous
chromosomes

Allele for white flowers

1-16
 Mendel’s Laws of Inheritance
• Homozygotes have two copies of the same
allele
• Heterozygotes have one copy of each allele
• Parents in 1st mating are homozygotes, having
2 copies of one allele
• Sex cells, or gametes, are haploid, containing
only 1 copy of each gene
• Heterozygotes produce gametes having either
allele
• Homozygotes produce gametes having only
one allele 1-17
 The Law of Independent
Assortment
• Mendel identified his second law of inheritance by
following two characters at the same time.

• Crossing two true-breeding parents differing in two

characters produces dihybrids in the F1 generation,
heterozygous for both characters.

• A dihybrid cross, a cross between F1 dihybrids, can

determine whether two characters are transmitted to
offspring as a package or independently.

1-18
 The Law of Independent
Assortment
• Using a dihybrid cross, Mendel developed the law of
independent assortment.

• The law of independent assortment states that each pair

of alleles segregates independently of each other pair of
alleles during gamete formation.

• Strictly speaking, this law applies only to genes on

different, nonhomologous chromosomes or those far
apart on the same chromosome.

• Genes located near each other on the same

chromosome tend to be inherited together. 1-19
 Summary
• Genes can exist in several different forms or
alleles
• One allele can be dominant over the other, so
heterozygotes having two different alleles of
one gene will generally exhibit the
characteristic dictated by the dominant allele
• The recessive allele is not lost; it can still exert
its influence when paired with another
recessive allele in a homozygote
1-20
 The Chromosome Theory of
Inheritance
• Chromosomes are discrete physical entities
that carry genes
• Thomas Hunt Morgan used the fruit fly,
Drosophila melanogaster, to study genetics
• Autosomes occur in pairs in a given individual
(not the X or the Y chromosome)
• Sex chromosomes are identified as X and Y
– Females have two X chromosomes
– Males have one X and one Y chromosome

1-21
Thomas Hunt Morgan

1-22
Morgan’s first mutant.

1-23
 Location of Genes on a
Chromosome
• Every gene has its place, or locus, on a
chromosome
• Genotype is the combination of alleles found in
an organism
• Phenotype is the visible expression of the
genotype
– Wild-type phenotype is the most common or
generally accepted standard
– Mutant alleles are usually recessive

1-24
 Linked genes tend to be inherited
together because they are located near
each other on the same chromosome
• Each chromosome has hundreds or thousands of genes
(except the Y chromosome).

• Genes located on the same chromosome that tend to be

inherited together are called linked genes.

• Morgan found that body color and wing size are usually
inherited together in specific combinations(parental
phenotypes).

• He noted that these genes do not assort independently,

and reasoned that they were on the same chromosome.
1-25
 How linkage affects inheritance

F1 dihybrid female b+ vg+ b vg

and homozygous
recessive male b vg b vg
in testcross

b+ vg+ b vg
Most offspring or
b vg b vg
1-26
Genetic Recombination and Mapping
• In early experiments genes on separate
chromosomes behaved independently

• Genes on the same chromosome behaved as if

they were linked

• This genetic linkage is not absolute

• Offspring show new combinations of alleles not

seen in the parents when recombination occurs
1-27
 Recombination
• During meiosis, gamete formation, crossing
over can occur resulting in the exchange of
genes between the two homologous
chromosomes

• The result of the crossing-over event produces

a new combination of alleles

• This process is called recombination

1-28
 Chromosomal basis for
recombination of linked genes.
Recombinant
chromosomes

bvg b vg b vg b vg
Eggs

Testcross 965 944 206 185

offspring Wild type Black- Gray- Black-
(gray-normal) vestigial vestigial normal b vg
b vg b vg b vg b vg

b vg b vg b vg b vg Sperm

Parental-type offspring Recombinant offspring

Recombination 391 recombinants  100  17%

frequency 
2,300 total offspring
1-29
 Genetic Mapping
• Morgan proposed that the farther apart two
genes are on a chromosome, the more likely
they are to recombine

• If two loci recombine with a frequency of 1%,

they are said to be separated by a map distance
of one centimorgan (named for Morgan)

• This mapping observation applies both to

prokaryotes and to eukaryotes
1-30
 Mapping the Distance
Between Genes
• Alfred Sturtevant, one of Morgan’s students,
constructed a genetic map, an ordered list of
the genetic loci along a particular
chromosome.

• Sturtevant predicted that the farther apart two

genes are, the higher the probability that a
crossover will occur between them and
therefore the higher the recombination
frequency.
1-31
 Constructing a Linkage Map

Recombination
frequencies
9% 9.5%
Chromosome
17%

b cn vg
Genes that are far apart on the same chromosome can have a
recombination frequency near 50%.

1-32
 Physical Evidence for
Recombination
• Microscopic examination of the maize
chromosome provided direct physical
observation of recombination using easily
identifiable features of one chromosome

• Similar observations were made in Drosophila

• Recombination was detected both physically

and genetically in both animals and plants

1-33
 Summary
• The chromosome theory of inheritance holds that genes
are arranged in linear fashion on chromosomes

• Certain traits tend to be inherited together when the

genes for those traits are on the same chromosome

• Recombination between two homologous

chromosomes during meiosis can scramble the
parental alleles to yield nonparental combinations

• The farther apart two genes are on a chromosome the

more likely it is that recombination will occur
1-34
 1.2 Molecular Genetics
• The Discovery of DNA: The general structure
of nucleic acids was discovered by the end of
the 19th century
– Long polymers or chains of nucleotides
– Nucleotides are linked by sugars through
phosphate groups
• Composition of Genes: DNA? RNA?
Protein? In 1944, Avery and his colleagues
demonstrated that genes are composed of DNA
1-35
 Evidence That DNA Can
Transform Bacteria
• The discovery of the genetic role of DNA began with
research by Frederick Griffith in 1928.

• Griffith worked with two strains of a bacterium, one

pathogenic and one harmless.

• When he mixed heat-killed remains of the pathogenic

strain with living cells of the harmless strain, some living
cells became pathogenic.

• He called this phenomenon transformation, now defined

as a change in genotype and phenotype due to
assimilation of foreign DNA. 1-36
Frederick Griffith

1-37
EXPERIMENT Mixture of
Heat-killed heat-killed
Living S cells Living R cells S cells S cells and
(control) (control) (control) living R cells

RESULTS

Mouse dies Mouse healthy Mouse healthy Mouse dies

Living S cells
1-38
 Evidence That DNA Can
Transform Bacteria
• In 1944, Oswald Avery, Maclyn McCarty, and Colin
MacLeod announced that the transforming substance
was DNA.

• Their conclusion was based on experimental evidence

that only DNA worked in transforming harmless bacteria
into pathogenic bacteria.

• Many biologists remained skeptical, mainly because

little was known about DNA.

1-39
Oswald Avery

Oswald Avery at work

in the laboratory,
around 1930 1-40
The experiments of Avery's lab
Illustration of the
classic
experiment by
Avery, MacLeod,
and McCarty
demonstrating
that DNA is
capable of
transforming
harmless R strain
S. pneumoniae
into the
pathogenic S
strain. 1-41
Evidence That Viral DNA Can
Program Cells
• More evidence for DNA as the genetic
material came from studies of viruses that
infect bacteria.

• Such viruses, called bacteriophages (or

phages), are widely used in molecular
genetics research.

1-42
Viruses infecting a bacterial cell
Phage
head

Tail
sheath
Tail fiber

DNA

100 nm
Bacterial
cell 1-43
 Alfred Hershey and Martha
Chase
• In 1952, Alfred Hershey and Martha Chase performed
experiments showing that DNA is the genetic material of
a phage known as T2.

• To determine this, they designed an experiment showing

that only one of the two components of T2 (DNA or
protein) enters an E. coli cell during infection.

• They concluded that the injected DNA of the phage

provides the genetic information.

1-44
 Radioactive Empty
protein protein
Radioactivity
Phage shell
(phage protein)
in liquid
Bacterial cell

Batch 1:
DNA
Radioactive
sulfur Phage
(35S) DNA

Centrifuge

Radioactive Pellet (bacterial

DNA cells and contents)

Batch 2:
Radioactive
phosphorus
(32P)
Centrifuge
Radioactivity
Pellet (phage DNA)
1-45
in pellet
 Additional Evidence That DNA
Is the Genetic Material
• It was known that DNA is a polymer of
nucleotides, each consisting of a sugar, a
nitrogenous base, and a phosphate group.

• In 1950, Erwin Chargaff reported that DNA

composition varies from one species to the next.

• This evidence of diversity made DNA a more

credible candidate for the genetic material.
1-46
 Additional Evidence That DNA
Is the Genetic Material
• Two findings became known as Chargaff’s rules
– The base composition of DNA varies between
species.

– In any species the number of A and T bases

are equal and the number of G and C bases
are equal.

• The basis for these rules was not understood

until the discovery of the double helix.
1-47
 Erwin Chargaff
In natural DNA, the number of
guanine units equals the
number of cytosine units and
the number of adenine units
equals the number of
thymine units.
In human DNA, for example,
the four bases are present in
these percentages: A=30.9%
and T=29.4%; G=19.9% and
C=19.8%.
The composition of DNA varies from one species to
another, in particular in the relative amounts of A, G, T,
and C bases. 1-48
 The double helix.
C G
5 end
C G Hydrogen bond
G C
3 end
G C T A

3.4 nm
T A
G C G C
C G
A T

1 nm T A
C G

C G
G C
C G A T

A T 3 end
A T
T A
0.34 nm 5 end
(a) Key features of (b) Partial chemical structure (c) Space-filling
DNA structure model 1-49
 The Relationship between Genes
and Proteins
• Experiments have shown that a defective gene
gives a defective or absent enzyme

• This lead to the proposal that one gene is

responsible for making one enzyme

• Proposal not quite correct for 3 reasons:

1. One enzyme may be composed of several
polypeptides, each gene codes for only one
polypeptide
2. Many genes code for non-enzyme proteins
3. End products of some genes are not polypeptides
(i.e. tRNA, rRNA) 1-50
 The Flow of Genetic
Information
• The information content of DNA is in the form of
specific sequences of nucleotides.

• The DNA inherited by an organism leads to specific

traits by dictating the synthesis of proteins.

• Proteins are the links between genotype and

phenotype.

• Gene expression, the process by which DNA directs

protein synthesis, includes two stages: transcription
and translation.
1-51
 Evidence from the Study of
Metabolic Defects
• In 1902, British physician Archibald Garrod first
suggested that genes dictate phenotypes through
enzymes that catalyze specific chemical reactions.

• He thought symptoms of an inherited disease reflect an

inability to synthesize a certain enzyme.

• Linking genes to enzymes required understanding that

cells synthesize and degrade molecules in a series of
steps, a metabolic pathway.

1-52
Sir Archibald Edward Garrod

1-53
 Nutritional Mutants in
Neurospora
• George Beadle and Edward Tatum exposed bread mold
to X-rays, creating mutants that were unable to survive
on minimal media.

• Using crosses, they and their coworkers identified three

classes of arginine-deficient mutants, each lacking a
different enzyme necessary for synthesizing arginine.

• They developed a one gene-one enzyme hypothesis,

which states that each gene dictates production of a
specific enzyme.

1-54
George Beadle and Edward
Tatum

1-55
George W. Beadle Edward L. Tatum
Neurospora Crassa

1-56
Do individual genes specify the enzymes
that function in a biochemical pathway?

EXPERIMENT

Growth: No growth:
Wild-type Mutant cells
cells growing cannot grow
and dividing and divide

Minimal medium

1-57
RESULTS
Classes of Neurospora crassa

Wild type Class I mutants Class II mutants Class III mutants

Minimal Growth
medium No
(MM) growth
(control)

MM 
ornithine
Condition

MM 
citrulline

MM 
arginine
(control)

Can grow with Can grow on Can grow only Require arginine
Summary ornithine,
or without any on citrulline or to grow
of results citrulline, or
supplements arginine
arginine 1-58
 Do individual genes specify the enzymes
that function in a biochemical pathway?
CONCLUSION
Gene Class I mutants Class II mutants Class III mutants
(codes for (mutation in (mutation in (mutation in
enzyme) Wild type gene A) gene B) gene C)

Precursor Precursor Precursor Precursor

Gene A Enzyme A Enzyme A Enzyme A Enzyme A

Ornithine Ornithine Ornithine Ornithine

Gene B Enzyme B Enzyme B Enzyme B Enzyme B

Citrulline Citrulline Citrulline Citrulline

Gene C Enzyme C Enzyme C Enzyme C Enzyme C

Arginine Arginine Arginine Arginine

1-59
Biosynthesis of Arginine in
Bacteria

1-60
 The Products of Gene
Expression: A Developing Story
• Some proteins aren’t enzymes, so researchers later
revised the hypothesis: one gene- one protein.

• Many proteins are composed of several polypeptides,

each of which has its own gene.

• Therefore, Beadle and Tatum’s hypothesis is now

restated as the one gene- one polypeptide hypothesis.

• Note that it is common to refer to gene products as

proteins rather than polypeptides.
1-61
 Activities of Genes
Genes perform three major roles
• Replicated faithfully
• Direct the production of RNAs and proteins
• Accumulate mutations thereby allowing for
evolution

DNA RNA Protein

1-62
 Replication
• Franklin and Wilkins produced x-ray diffraction
data on DNA, Watson and Crick proposed that
DNA is double helix
– Two DNA strands wound around each other
– Strands are complementary – if you know the
sequence of one strand, you automatically know the
sequence of the other strand

• Semiconservative replication keeps one strand

of the parental double helix conserved in each
of the daughter double helices
1-63
 Building a Structural Model of
DNA
• After DNA was accepted as the genetic material,
the challenge was to determine how its structure
accounts for its role in heredity.

• Maurice Wilkins and Rosalind Franklin were

using a technique (X-ray crystallography) to
study molecular structure.

• Franklin produced a picture of the DNA molecule

using this technique.
1-64
Rosalind Franklin and her X-
ray diffraction photo of DNA.

(a) Rosalind Franklin (b) Franklin’s X-ray diffraction

photograph of DNA 1-65
 Watson and Crick
• Watson and Crick built models of a double helix
to conform to the X-rays and chemistry of DNA.

• Franklin had concluded that there were two

outer sugar-phosphate backbones, with the
nitrogenous bases paired in the molecule’s
interior.

• Watson built a model in which the backbones

were antiparallel (their subunits run in opposite
directions).
1-66
 A model for DNA replication:
the basic concept.
A T A T A T A T
C G C G C G C G
T A T A T A T A
A T A T A T A T
G C G C G C G C

(a) Parent molecule (b) Separation of (c)“Daughter” DNA molecules,

strands each consisting of one
parental strand and one
new strand

1-67
 Semiconservative model
• Watson and Crick’s semiconservative model of
replication predicts that when a double helix
replicates, each daughter molecule will have one
old strand (derived or “conserved” from the
parent molecule) and one newly made strand.

• Competing models were the conservative model

(the two parent strands rejoin) and the
dispersive model(each strand is a mix of old and
new).
1-68
 Parent First Second
cell replication replication
(a) Conservative
model

(b) Semiconservative
model

Three
(c) Dispersive model
alternative
models of DNA
replication.
1-69
 Semiconservative model
• Experiments by Matthew Meselson and Franklin Stahl
supported the semiconservative model.

• They labeled the nucleotides of the old strands with a

heavy isotope of nitrogen, while any new nucleotides
were labeled with a lighter isotope.

• The first replication produced a band of hybrid DNA,

eliminating the conservative model.

• A second replication produced both light and hybrid

DNA, eliminating the dispersive model and supporting
the semiconservative model.
1-70
Matthew Meselson and
Franklin Stahl

1-71
Does DNA replication follow the conservative,
semiconservative, or dispersive model?
EXPERIMENT
1 Bacteria 2 Bacteria
cultured in transferred to
medium with medium with
15N (heavy 14N (lighter

isotope) isotope)

RESULTS
3 DNA sample 4 DNA sample Less
centrifuged centrifuged dense
after first after second
replication replication More
dense
1-72
 Does DNA replication follow the
CONCLUSION
Predictions: First replication
conservative, Second replication
semiconservative, or
dispersive model?
Conservative
model

Semiconservative
model

Dispersive
model
1-73
 Genes Direct the Production of
Polypeptides
• Gene expression is the process by which a
gene product is made

• Two steps are required

– 1. Transcription: DNA is transcribed into RNA
– 2. Translation: the mRNA is read or translated to
assemble a protein
– Codon: a sequence of 3 nucleic acid bases that code
for one amino acid within the mRNA

1-74
 The Genetic Code
• How are the instructions for assembling
amino acids into proteins encoded into DNA?

• There are 20 amino acids, but there are only

four nucleotide bases in DNA.

• How many nucleotides correspond to an

amino acid?

1-75
Codons: Triplets of Nucleotides
• The flow of information from gene to protein is
based on a triplet code: a series of
nonoverlapping, three-nucleotide words.

• The words of a gene are transcribed into

complementary nonoverlapping three-nucleotide
words of mRNA.

• These words are then translated into a chain of

amino acids, forming a polypeptide.
1-76
 Genes Accumulate Mutations
Genes change in several ways
• Change one base to another
• Deletions of one base up to a large segment
• Insertions of one base up to a large segment
• The more drastic the change, the more likely it
is that the gene or genes involved will be totally
inactivated

1-77
 Mutations of one or a few nucleotides
can affect protein structure and function
• Mutations are changes in the genetic material of
a cell or virus.

• Point mutations are chemical changes in just

one base pair of a gene.

• The change of a single nucleotide in a DNA

template strand can lead to the production of an
abnormal protein.
1-78
Types of Small-Scale Mutations
• Point mutations within a gene can be divided into two
general categories

– Nucleotide-pair substitutions

– One or more nucleotide-pair insertions or

deletions

1-79
 Substitutions
• A nucleotide-pair substitution replaces one nucleotide
and its partner with another pair of nucleotides

• Silent mutations have no effect on the amino acid

produced by a codon because of redundancy in the
genetic code

• Missense mutations still code for an amino acid, but not

the correct amino acid

• Nonsense mutations change an amino acid codon into a

stop codon, nearly always leading to a nonfunctional
protein
1-80
Types of small-scale mutations
that affect mRNA sequence.
Wild type
DNA template strand 3 T A C T T C A A A C C G A T T 5
5 A T G A A G T T T G G C T A A 3

mRNA5 A U G A A G U U U G G C U A A 3
Protein Met Lys Phe Gly Stop
Amino end Carboxyl end

(a) Nucleotide-pair substitution: silent

A instead of G
3 T A C T T C A A A C C A A T T 5
5 A T G A A G T T T G G T T A A 3
U instead of C
5 A U G A A G U U U G G U U A A 3
1-81
Met Lys Phe Gly Stop
Types of small-scale mutations
that affect mRNA sequence.
Wild type
DNA template strand 3 T A C T T C A A A C C G A T T 5
5 A T G A A G T T T G G C T A A 3

mRNA5 A U G A A G U U U G G C U A A 3
Protein Met Lys Phe Gly Stop
Amino end Carboxyl end

(a) Nucleotide-pair substitution: missense

T instead of C
3 T A C T T C A A A T C G A T T 5
5 A T G A A G T T T A G C T A A 3
A instead of G
5 A U G A A G U U U A G C U A A 3
1-82
Met Lys Phe Ser Stop
Types of small-scale mutations
that affect mRNA sequence.
Wild type
DNA template strand 3 T A C T T C A A A C C G A T T 5
5 A T G A A G T T T G G C T A A 3

mRNA5 A U G A A G U U U G G C U A A 3
Protein Met Lys Phe Gly Stop
Amino end Carboxyl end

(a) Nucleotide-pair substitution: nonsense

A instead of T T instead of C
3 T A C A T C A A A C C G A T T 5
5 A T G T A G T T T G G C T A A 3
U instead of A
5 A U G U A G U U U G G C U A A 3
1-83
Met Stop
 Insertions and Deletions
• Insertions and deletions are additions or losses
of nucleotide pairs in a gene

• These mutations have a disastrous effect on the

resulting protein more often than substitutions
do

• Insertion or deletion of nucleotides may alter the

reading frame, producing a frameshift mutation

1-84
Types of small-scale mutations
that affect mRNA sequence.
Wild type
DNA template strand 3 T A C T T C A A A C C G A T T 5
5 A T G A A G T T T G G C T A A 3
mRNA5 A U G A A G U U U G G C U A A 3
Protein Met Lys Phe Gly Stop
Amino end Carboxyl end

(b) Nucleotide-pair insertion or deletion: frameshift causing

immediate nonsense
Extra A
3 T A C A T T C A A A C G G A T T 5
5 A T G T A A G T T T G G C T A A 3
Extra U
5 A U G U A A G U U U G G C U A A 3
Met Stop 1-85
1 nucleotide-pair insertion
Types of small-scale mutations
that affect mRNA sequence.
Wild type
DNA template strand 3 T A C T T C A A A C C G A T T 5
5 A T G A A G T T T G G C T A A 3
mRNA5 A U G A A G U U U G G C U A A 3
Protein Met Lys Phe Gly Stop
Amino end Carboxyl end

(b) Nucleotide-pair insertion or deletion: frameshift causing

extensive missense
A missing
3 T A C T T C A A C C G A T T 5
5 A T G A A G T T G G C T A A 3
U missing
5 A U G A A G U U G G C U A A 3
Met Lys Leu Ala 1-86
1 nucleotide-pair deletion
Types of small-scale mutations
that affect mRNA sequence.
Wild type
DNA template strand 3 T A C T T C A A A C C G A T T 5
5 A T G A A G T T T G G C T A A 3
mRNA5 A U G A A G U U U G G C U A A 3
Protein Met Lys Phe Gly Stop
Amino end Carboxyl end

(b) Nucleotide-pair insertion or deletion: no frameshift, but one

amino acid missing
T T C missing
3 T A C A A A C C G A T T 5
5 A T G T T T G G C T A A 3
A A G missing
5 A U G U U U G G C U A A 3
Met Phe Gly Stop 1-87
3 nucleotide-pair deletion
 Summary
• All cellular genes are made of DNA arranged in
a double helix

• This structure explains how genes replicate,

carry information and collect mutations

• The sequence of nucleotides in a gene is a

genetic code that carries the information for
making an RNA

• A change in the sequence of bases

constitutes and mutation, which can change
the sequence of amino acids in the genes
polypeptide product 1-88
 1.3 The Three Domains of Life
Current research theories support the
division of living organisms into three
domains
1. Bacteria
2. Eukaryota
3. Archaea
• Like bacteria as they are organisms without nuclei
• More similar to eukaryotes in the context of their
molecular biology

1-89
Kingdoms of life

1-90
 The three domains of Life
(a) Domain Bacteria (b) Domain Archaea

2 m
2 m
(c) Domain Eukarya
Kingdom Animalia

100 m

Kingdom Plantae
Protists

Kingdom Fungi 1-91

Bacteria
Eubacteria literally means true
bacteria

Prokaryotic organisms

Cell walls made of

peptidoglycan

Predominantly unicellular.

Microscopic

E. coli, Staphylococcus,
Salmonella, Lactobacillus,1-92
etc.
Archaea
Prokaryotic organisms

Most Archaea are anaerobic

Extremophiles

Predominantly unicellular.

Microscopic

About 250 species have been

identified
1-93
 Archaea
Archaea live in the most inhospitable regions of
the earth
• Thermophiles tolerate extremely high
temperatures
• Halophiles tolerate very high salt
concentrations
• Methanogens produce methane as a by-
product of metabolism

1-94
 Difference between bacteria
and archaea
1) The base "thymine" is not present in tRNA of archaea
2) The first amino acid is methionine in archaea (it is not
N-formylated)
3) Promoter structures are different
5) Structure of their ATPases are different
6) Methanogenesis is unique to archaea
7) Some archaea are photosynthetic, and it is strictly non-
chlorophyll based. Photosynthesis in bacteria (and
eukaryotes) is strictly chlorophyll based.

1-95
 Eukarya
• Protista Kingdom: Protista are simple, usually unicellular eukaryotic
organisms. Examples include slime molds (fungi-like protists), algae
(plant-like protists), and protozoans (animal-like protists).
• Fungi Kingdom: Fungi are typically multicellular absorptive
heterotrophs. (They digest their food before they ingest it) Their cells have
cell walls made of chitin but are not organized into tissues. They do not
carry out photosynthesis and obtain nutrients through absorption.
Examples include sac fungi, club fungi, yeasts, mushroom and molds.
• Animalia Kingdom: Animals are ingestive (they ingest their food
before they digest it) heterotrphic multicellular organisms. The cells are
organized into tissues and lack cell walls. They do not carry out
photosynthesis and obtain nutrients primarily by ingestion. Examples
include sponges, worms, insects, and vertebrates.
• Plantae Kingdom: Plants are autotrophic multicellular organisms
composed of eukaryotic cells. The cells are organized into tissues and
have cell walls of cellulose. They obtain nutrients by photosynthesis and
absorption. Examples include mosses, ferns, conifers, and flowering 1-96
plants.