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UNIT 1:
GENETICS:
NUCLEIC ACID
DNA
(Campbell & Reece:
Chapters, 5, 16)
LOCATION OF DNA IN
A CELL

• Chromatin is a complex of DNA and
protein, and is found in the nucleus of
eukaryotic cells.
• Histones are proteins that are
responsible for the first level of DNA
packing in chromatin
• The chromatin network in the nucleus of
a cell will coil up tightly during cell
division and form individual
chromosomes.
• Chromosomes are always duplicated
during this process (2 sets of identical
genetic information to ensure each cell
receives identical genetic info to the
parent cell during cell division).
• A duplicated chromosome consists of 2
chromatids attached to each other by a
centromere.
• Each chromatid consists of several genes.
• Genes consists of a long DNA strand.
• A string of DNA coiled around a few
histones is called a nucleosome.
LOCATION OF DNA IN
A CELL

Locus: Position of gene on chromosome
CHROMOSOME DUPLICATION
AND DISTRIBUTION
2. DNA STRUCTURE
• DNA molecules are polymers called
polynucleotides.
• Each polynucleotide is made of
monomers called nucleotides.
• Each nucleotide consists of :
• a nitrogenous base
(Adenine, Thymine, Cytosine or
Guanine)
• a pentose sugar (DNA =
Deoxyribose sugar),
• Nucleotide monomers are linked
together to build a polynucleotide.
• Adjacent nucleotides are joined by
covalent bonds that form between the –
OH group on the 3’ carbon of one
nucleotide and the phosphate on the 5’
carbon on the next nucleotide.
• These links create a backbone of sugarphosphate units with nitrogenous bases
as appendages.
• The sequence of bases along a DNA
polymer is unique for each gene.
A polynucleotide and a single nucleotide
The different nitrogenous bases in DNA
Single ring structure

Double ring structure
• A DNA molecule has two polynucleotides spiralling around an imaginary
axis, forming a double helix.
• In the DNA double helix, the two
backbones run in opposite 5’ → 3’
directions from each other, an
arrangement referred to as antiparallel.
• One DNA molecule includes many genes.
• The nitrogenous bases in DNA pair up
and form hydrogen bonds: adenine (A)
always with thymine (T), and guanine
(G) always with cytosine (C).
A DNA double helix structure
3. DISCOVERY OF THE
DNA STRUCTURE

• Early in the 20th century, the identification of the molecules of inheritance
loomed as a major challenge to biologists.
The Search for the Genetic
Material: Scientific Inquiry
• T. H. Morgan’s group showed that genes
are located on chromosomes, the 2
components of chromosomes are DNA &
protein which became the candidates for
the genetic material.
• Key factor in determining the genetic
material was choosing appropriate
experimental organisms - bacteria & the
viruses that infect them were chosen.
• Discovery of the genetic role of DNA began
with research by Frederick Griffith in 1928.
• Griffith worked with 2 strains of a
bacterium, 1 pathogenic (S cells) & 1
harmless (R cells)
• Heat-killed pathogenic strain were mixed
with living cells of harmless strain and the
result = some living cells became
pathogenic.
• This phenomenon was called
transformation, now defined as a change
in genotype & phenotype due to
assimilation of foreign DNA.
• 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.
Phage head

Tail sheath

Tail

fiber

DNA

Bacterial cell
• 1952: A. Hershey & M. Chase
experiments showing that DNA is
the genetic material of T2 phage.
• To determine the source of genetic
material in the phage, they designed
an experiment showing that only 1 /
2 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.
• After most biologists became
convinced that DNA was the genetic
material, the challenge was to
determine how its structure accounts
for its role…
• M Wilkins & R Franklin used X-ray
crystallography to study molecular
structure.
• Franklin produced a picture of the DNA
molecule using this technique.
A picture of the DNA molecule using
crystallography by Franklin.
• Franklin’s X-ray crystallographic
images of DNA enabled Watson to
deduce:
• that DNA was helical
• the width of the helix
• the spacing of the nitrogenous
bases
• Width suggested that the DNA
molecule was made up of 2
strands, forming a double helix
Representations of DNA molecule
• Watson and Crick: built models of a
double helix to conform to the X-rays &
chemistry of DNA.
• Franklin concluded there were 2
antiparallel sugar-P backbones, with
the N bases paired in the molecule’s
interior.
• But: How did bases pair? A-A?/A-T?/
A-C?/A-G?......
• But: How did bases pair?
• They worked it out by using the
following image:
Purine + purine: too wide

Pyrimidine + pyrimidine:
too narrow
Purine + pyrimidine: width
consistent with X-ray data
• W & C noted that the pairing of the
Nitrogen bases was specific, dictated
by the base structures.
• Adenine (A) paired only with thymine
(T), & guanine (G) paired only with
cytosine (C)
• The W-C model explains Chargaff’s
rules which states that; in any
organism the amount of A = T, & the
amount of G = C
4. THE ROLE OF DNA
• DNA is vital for all living beings – even
plants.
• It is important for:
• inheritance,
• coding for proteins and
• the genetic instruction guide for
life and its processes.
 DNA holds the instructions for an
organism's or each cell’s development
and reproduction and ultimately death.
 DNA can replicate itself.
NON-CODING DNA
 Multicellular eukaryotes have many
introns(non-coding DNA) within genes and
noncoding DNA between genes.
 The bulk of most eukaryotic genomes
consists of noncoding DNA
sequences, often described in the past as
“junk DNA”
 Much evidence indicates that noncoding
DNA plays important roles in the cell.
 Sequencing of the human genome reveals
that 98.5% does not code for
proteins, rRNAs, or tRNAs.
 About 24% of the human genome
codes for introns and gene-related
regulatory sequences.
 Intergenic DNA is noncoding DNA
found between genes:
 Pseudogenes are former genes that
have accumulated mutations and are
nonfunctional
 Repetitive DNA is present in
multiple copies in the genome
5. DNA REPLICATION

• Replication begins at special sites
called origins of replication, where
the 2 DNA strands separate, opening
up a replication “bubble” (eukaryotic
chromosome may have many origins
of replication.
• The enzyme helicase unwinds the
parental double helix.
• Single stranded binding protein
stabilizes the unwound template
strands.
• A replication fork forms.
• The enzyme: Topoisomerase
breaks, swivels and re-joins the
parental DNA ahead of the replication
for, to prevent over winding.
• The unwind complimentary strands
now act as individual template for 2
new strands.
• RNA nucleotides are added to each
DNA template by the enzyme RNA
primase to form RNA primers on both
templates
• The enzyme DNA polymerase III add
free DNA nucleotides to the RNA
primer 3’ carbon.
• The free nucleotides bond with Hbonds to their complimentary bases
on the DNA templates.
• Along one template strand of DNA,
the DNA polymerase synthesizes a
leading strand continuously, moving
toward the replication fork.
• To elongate the other new strand,
called the lagging strand, DNA
polymerase must work in the
direction away from the replication
fork.
• The lagging strand is synthesized as a
series of segments called Okazaki
fragments.
• Each fragment has an RNA primer and
added DNA strand.
• All the fragments are then joined with
the help of the enzyme DNA ligase.
• DNA polymerase I then removes RNA
primers and replaces it with DNA
nucleotides.
6. PROOFREADING AND
REPAIRING DNA
• DNA polymerases proofread newly made
DNA, replacing any incorrect nucleotides
• In mismatch repair of DNA, repair
enzymes correct errors in base pairing.
• DNA damaged by chemicals, radioactive
emissions, X-rays, UV light, & certain
molecules (in cigarette smoke for example)
• In nucleotide excision repair, a nuclease
cuts out & replaces damaged stretches of
DNA.
7.BIOTECHNOLOGY
AND GENOMICS
DNA CLONING
BIOTECH PRODUCTS
DNA CLONING
–Cloning is the reproduction of genetically
identical copies of DNA, cells or
organisms through some asexual means.
–DNA cloning can be done to produce
many identical copies of the same gene –
for the purpose of gene cloning.
–When cloned genes are used to modify a
human, the process is called gene
therapy.
–Otherwise, the organisms are called
transgenic organisms – these
organisms today are used to produce
products desired by humans.
–A. Recombinant DNA (rDNA) and B.
polymerase chain reaction (PCR) are
two procedures that scientists can use
to clone DNA
CLONING OF AN ORGANISM
A. CLONING A HUMAN GENE
HOW IS INSULIN MADE BY DNA CLONING?
(A. RECOMBINANT DNA)
• A large quantity of insulin are being produced
by recombinant DNA technology.
• This process is as follows:
1. DNA that codes for the production of
insulin is removed from the chromosome of a
human pancreatic cell.
2. Restriction enzymes cut the gene from the
chromosome (isolating the gene for insulin)
I
Insulin gene – cut
out with restriction
enzyme
Isolated insulin gene
3. A plasmid (acting as a vector/carrier of

new gene) is removed from the bacterium
and cut open with a restriction enzyme to
form sticky ends
Nucleus

plasmid

BACTERIUM CELL

Sticky ends

Plasmid removed from
bacterium

Cut by restriction
enzyme
• 4. Ligase (enzyme) is added to join the
insulin gene to the plasmid of the
bacterium cell - forming recombinant
DNA.
Insulin gene placed in plasmid
by enzyme Ligase ( attached
to sticky ends)

The recombinant DNA can then be
reinserted into the bacterium, the
bacterium will then produce more
insulin, therefore cloning the gene.

• 5.
Recombinant DNA placed into bacterium cell
• 6. When the bacterium reproduces it makes
the insulin inserted into the plasmid.
• 7. The bacteria are kept in huge tenks with
optimum pH, temperature and nutrient
values, where they multiply rapidly, producing
enormous amounts of insulin, this is then
purified and sold.
B. POLYMERASE CHAIN REACTION
• PCR – Used in genetic profiling.
• To solve crimes – criminals usually leave DNA
evidence at the scene of the crime in the form
of saliva, blood, skin, semen and hair. These all
contain DNA. If only a little bit of DNA is found
or the DNA is old, we can make copies of the
available DNA by means of PCR.
• From the DNA produced through PCR, DNA
fingerprint can be generated.
PCR method
• 1. Sample containing DNA is heated in a
test tube to separate DNA into single
strands.
• 2. Free nucleotides are added to the test
tube with DNA polymerase (enzyme), to
allow DNA replication.
• 3. DNA is cooled to allow free nucleotides
to form a complementary strand along side
each single strand.
• 4. In this way the DNA is doubled giving
sufficient amount of DNA to work with.
DNA SCREENING AND FINGERPRINT
TECHNIQUE

• 1. Sample of DNA is cut into fragments by means of
restriction enzymes.
• 2. Negative charged electrode at one end of a
rectangular flat piece of gel and a positive
electrode is placed at the other end.
• 3. The DNA is placed at the negative end of the gel
and starts to move to the positive end. Smaller
fragments move faster than the larger ones.
Separation occurs on the basis of size. This process
is called gel electrophoresis.
• 4. DNA is then pressed flat against the gel and
transferred to filter paper.
• 5. Radioactive probes bind to special DNA
fragments.
• 6. X-rays are taken of the filter paper. The DNA
probes show up as dark bands on the film. The
pattern of these bands is the DNA fingerprint.
DNA fingerprinting
GEL ELECTROPHORESIS
DNA FINGERPRINT
BIOTECHNOLOGY PRODUCTS
• Today transgenic bacteria, plants and animals
are called genetically modified organisms
(GMO’s).
• The products that GMO’s produce are called
biotechnology products.
GENETICALLY MODIFIED BACTERIA
• Recombinant DNA is used to make transgenic
bacteria.
• They are used to make insulin, clotting factor
VIII, human growth hormone and hepatitis B
vaccine.
• Transgenic bacteria is used to protect the
roots of plants from insect attack, by
producing insect toxins.
GENETICALLY MODIFIED PLANTS
• Example = pomato
• Genetically modified to produce potato's
below the ground and tomato's above the
ground.
• Foreign genes transferred to cotton, corn, and
potato strains have made these plants
resistant to pests because their cells now
produce an insect toxin.
• Read p. 253 for more examples
GENETICALLY MODIFIED ANIMALS

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Unit 1 genetics nucleic acids dna

  • 2. LOCATION OF DNA IN A CELL • Chromatin is a complex of DNA and protein, and is found in the nucleus of eukaryotic cells. • Histones are proteins that are responsible for the first level of DNA packing in chromatin • The chromatin network in the nucleus of a cell will coil up tightly during cell division and form individual chromosomes.
  • 3. • Chromosomes are always duplicated during this process (2 sets of identical genetic information to ensure each cell receives identical genetic info to the parent cell during cell division). • A duplicated chromosome consists of 2 chromatids attached to each other by a centromere. • Each chromatid consists of several genes. • Genes consists of a long DNA strand. • A string of DNA coiled around a few histones is called a nucleosome.
  • 4. LOCATION OF DNA IN A CELL Locus: Position of gene on chromosome
  • 6. 2. DNA STRUCTURE • DNA molecules are polymers called polynucleotides. • Each polynucleotide is made of monomers called nucleotides. • Each nucleotide consists of : • a nitrogenous base (Adenine, Thymine, Cytosine or Guanine) • a pentose sugar (DNA = Deoxyribose sugar),
  • 7. • Nucleotide monomers are linked together to build a polynucleotide. • Adjacent nucleotides are joined by covalent bonds that form between the – OH group on the 3’ carbon of one nucleotide and the phosphate on the 5’ carbon on the next nucleotide. • These links create a backbone of sugarphosphate units with nitrogenous bases as appendages. • The sequence of bases along a DNA polymer is unique for each gene.
  • 8. A polynucleotide and a single nucleotide
  • 9. The different nitrogenous bases in DNA Single ring structure Double ring structure
  • 10. • A DNA molecule has two polynucleotides spiralling around an imaginary axis, forming a double helix. • In the DNA double helix, the two backbones run in opposite 5’ → 3’ directions from each other, an arrangement referred to as antiparallel. • One DNA molecule includes many genes. • The nitrogenous bases in DNA pair up and form hydrogen bonds: adenine (A) always with thymine (T), and guanine (G) always with cytosine (C).
  • 11. A DNA double helix structure
  • 12. 3. DISCOVERY OF THE DNA STRUCTURE • Early in the 20th century, the identification of the molecules of inheritance loomed as a major challenge to biologists.
  • 13. The Search for the Genetic Material: Scientific Inquiry • T. H. Morgan’s group showed that genes are located on chromosomes, the 2 components of chromosomes are DNA & protein which became the candidates for the genetic material. • Key factor in determining the genetic material was choosing appropriate experimental organisms - bacteria & the viruses that infect them were chosen.
  • 14. • Discovery of the genetic role of DNA began with research by Frederick Griffith in 1928. • Griffith worked with 2 strains of a bacterium, 1 pathogenic (S cells) & 1 harmless (R cells) • Heat-killed pathogenic strain were mixed with living cells of harmless strain and the result = some living cells became pathogenic. • This phenomenon was called transformation, now defined as a change in genotype & phenotype due to assimilation of foreign DNA.
  • 15.
  • 16. • 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. Phage head Tail sheath Tail fiber DNA Bacterial cell
  • 17. • 1952: A. Hershey & M. Chase experiments showing that DNA is the genetic material of T2 phage. • To determine the source of genetic material in the phage, they designed an experiment showing that only 1 / 2 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.
  • 18.
  • 19. • After most biologists became convinced that DNA was the genetic material, the challenge was to determine how its structure accounts for its role… • M Wilkins & R Franklin used X-ray crystallography to study molecular structure. • Franklin produced a picture of the DNA molecule using this technique.
  • 20. A picture of the DNA molecule using crystallography by Franklin.
  • 21. • Franklin’s X-ray crystallographic images of DNA enabled Watson to deduce: • that DNA was helical • the width of the helix • the spacing of the nitrogenous bases • Width suggested that the DNA molecule was made up of 2 strands, forming a double helix
  • 23. • Watson and Crick: built models of a double helix to conform to the X-rays & chemistry of DNA. • Franklin concluded there were 2 antiparallel sugar-P backbones, with the N bases paired in the molecule’s interior. • But: How did bases pair? A-A?/A-T?/ A-C?/A-G?......
  • 24. • But: How did bases pair? • They worked it out by using the following image: Purine + purine: too wide Pyrimidine + pyrimidine: too narrow Purine + pyrimidine: width consistent with X-ray data
  • 25. • W & C noted that the pairing of the Nitrogen bases was specific, dictated by the base structures. • Adenine (A) paired only with thymine (T), & guanine (G) paired only with cytosine (C) • The W-C model explains Chargaff’s rules which states that; in any organism the amount of A = T, & the amount of G = C
  • 26.
  • 27. 4. THE ROLE OF DNA • DNA is vital for all living beings – even plants. • It is important for: • inheritance, • coding for proteins and • the genetic instruction guide for life and its processes.  DNA holds the instructions for an organism's or each cell’s development and reproduction and ultimately death.  DNA can replicate itself.
  • 28. NON-CODING DNA  Multicellular eukaryotes have many introns(non-coding DNA) within genes and noncoding DNA between genes.  The bulk of most eukaryotic genomes consists of noncoding DNA sequences, often described in the past as “junk DNA”  Much evidence indicates that noncoding DNA plays important roles in the cell.  Sequencing of the human genome reveals that 98.5% does not code for proteins, rRNAs, or tRNAs.
  • 29.  About 24% of the human genome codes for introns and gene-related regulatory sequences.  Intergenic DNA is noncoding DNA found between genes:  Pseudogenes are former genes that have accumulated mutations and are nonfunctional  Repetitive DNA is present in multiple copies in the genome
  • 30. 5. DNA REPLICATION • Replication begins at special sites called origins of replication, where the 2 DNA strands separate, opening up a replication “bubble” (eukaryotic chromosome may have many origins of replication. • The enzyme helicase unwinds the parental double helix. • Single stranded binding protein stabilizes the unwound template strands.
  • 31. • A replication fork forms. • The enzyme: Topoisomerase breaks, swivels and re-joins the parental DNA ahead of the replication for, to prevent over winding. • The unwind complimentary strands now act as individual template for 2 new strands. • RNA nucleotides are added to each DNA template by the enzyme RNA primase to form RNA primers on both templates
  • 32.
  • 33. • The enzyme DNA polymerase III add free DNA nucleotides to the RNA primer 3’ carbon. • The free nucleotides bond with Hbonds to their complimentary bases on the DNA templates. • Along one template strand of DNA, the DNA polymerase synthesizes a leading strand continuously, moving toward the replication fork.
  • 34.
  • 35. • To elongate the other new strand, called the lagging strand, DNA polymerase must work in the direction away from the replication fork. • The lagging strand is synthesized as a series of segments called Okazaki fragments. • Each fragment has an RNA primer and added DNA strand. • All the fragments are then joined with the help of the enzyme DNA ligase.
  • 36. • DNA polymerase I then removes RNA primers and replaces it with DNA nucleotides.
  • 37. 6. PROOFREADING AND REPAIRING DNA • DNA polymerases proofread newly made DNA, replacing any incorrect nucleotides • In mismatch repair of DNA, repair enzymes correct errors in base pairing. • DNA damaged by chemicals, radioactive emissions, X-rays, UV light, & certain molecules (in cigarette smoke for example) • In nucleotide excision repair, a nuclease cuts out & replaces damaged stretches of DNA.
  • 38.
  • 40. DNA CLONING –Cloning is the reproduction of genetically identical copies of DNA, cells or organisms through some asexual means. –DNA cloning can be done to produce many identical copies of the same gene – for the purpose of gene cloning. –When cloned genes are used to modify a human, the process is called gene therapy.
  • 41. –Otherwise, the organisms are called transgenic organisms – these organisms today are used to produce products desired by humans. –A. Recombinant DNA (rDNA) and B. polymerase chain reaction (PCR) are two procedures that scientists can use to clone DNA
  • 42. CLONING OF AN ORGANISM
  • 43. A. CLONING A HUMAN GENE HOW IS INSULIN MADE BY DNA CLONING? (A. RECOMBINANT DNA) • A large quantity of insulin are being produced by recombinant DNA technology. • This process is as follows: 1. DNA that codes for the production of insulin is removed from the chromosome of a human pancreatic cell. 2. Restriction enzymes cut the gene from the chromosome (isolating the gene for insulin) I
  • 44. Insulin gene – cut out with restriction enzyme Isolated insulin gene
  • 45. 3. A plasmid (acting as a vector/carrier of new gene) is removed from the bacterium and cut open with a restriction enzyme to form sticky ends Nucleus plasmid BACTERIUM CELL Sticky ends Plasmid removed from bacterium Cut by restriction enzyme
  • 46. • 4. Ligase (enzyme) is added to join the insulin gene to the plasmid of the bacterium cell - forming recombinant DNA. Insulin gene placed in plasmid by enzyme Ligase ( attached to sticky ends) The recombinant DNA can then be reinserted into the bacterium, the bacterium will then produce more insulin, therefore cloning the gene. • 5.
  • 47. Recombinant DNA placed into bacterium cell • 6. When the bacterium reproduces it makes the insulin inserted into the plasmid. • 7. The bacteria are kept in huge tenks with optimum pH, temperature and nutrient values, where they multiply rapidly, producing enormous amounts of insulin, this is then purified and sold.
  • 48. B. POLYMERASE CHAIN REACTION • PCR – Used in genetic profiling. • To solve crimes – criminals usually leave DNA evidence at the scene of the crime in the form of saliva, blood, skin, semen and hair. These all contain DNA. If only a little bit of DNA is found or the DNA is old, we can make copies of the available DNA by means of PCR. • From the DNA produced through PCR, DNA fingerprint can be generated.
  • 49. PCR method • 1. Sample containing DNA is heated in a test tube to separate DNA into single strands. • 2. Free nucleotides are added to the test tube with DNA polymerase (enzyme), to allow DNA replication. • 3. DNA is cooled to allow free nucleotides to form a complementary strand along side each single strand. • 4. In this way the DNA is doubled giving sufficient amount of DNA to work with.
  • 50.
  • 51.
  • 52. DNA SCREENING AND FINGERPRINT TECHNIQUE • 1. Sample of DNA is cut into fragments by means of restriction enzymes. • 2. Negative charged electrode at one end of a rectangular flat piece of gel and a positive electrode is placed at the other end. • 3. The DNA is placed at the negative end of the gel and starts to move to the positive end. Smaller fragments move faster than the larger ones. Separation occurs on the basis of size. This process is called gel electrophoresis.
  • 53. • 4. DNA is then pressed flat against the gel and transferred to filter paper. • 5. Radioactive probes bind to special DNA fragments. • 6. X-rays are taken of the filter paper. The DNA probes show up as dark bands on the film. The pattern of these bands is the DNA fingerprint.
  • 57. BIOTECHNOLOGY PRODUCTS • Today transgenic bacteria, plants and animals are called genetically modified organisms (GMO’s). • The products that GMO’s produce are called biotechnology products.
  • 58.
  • 59. GENETICALLY MODIFIED BACTERIA • Recombinant DNA is used to make transgenic bacteria. • They are used to make insulin, clotting factor VIII, human growth hormone and hepatitis B vaccine. • Transgenic bacteria is used to protect the roots of plants from insect attack, by producing insect toxins.
  • 60. GENETICALLY MODIFIED PLANTS • Example = pomato • Genetically modified to produce potato's below the ground and tomato's above the ground. • Foreign genes transferred to cotton, corn, and potato strains have made these plants resistant to pests because their cells now produce an insect toxin. • Read p. 253 for more examples