CAMPBELL BIOLOGY: CONCEPTS & CONNECTIONS
Tenth
Edition
CHAPTER 12
DNA TECHNOLOGY AND
GENOMICS
BIOL 1306 – General Biology 1
Ethan Ebner, M.S.
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Gene Cloning and Editing
DNA technology – methods used to study/manipulate
DNA
Recombinant DNA – a DNA molecule that has been
manipulated to carry genetic DNA from two different
sources/species
Plasmids – a small, circular independently replicating
DNA sequence found in prokaryotes, a key tool for
DNA cloning
Carry only a few genes
Easily transferred from bacteria to bacteria
Passed from one generation to the next rapidly
Easily manipulated
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Gene Cloning and Editing
Vector – a piece of DNA that is used to move genes
from one cell to another, often a bacterial plasmid
Gene cloning – the production of multiple copies of a
gene
1. The gene is inserted into another organism
directly, new organism can now utilize that gene
2. The protein product of the gene is harvested in
large quantities
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Gene Cloning
and Editing
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Gene Cloning and Editing
Restriction Enzymes – bacterial enzymes that cut
up foreign DNA
Restriction Site – the “cut site” for a restriction
enzyme, the specific sequence a restriction
enzyme works at
Restriction Fragments – the yielding pieces after
a restriction enzyme has cut, cuts DNA at precise
points within a sequence
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Gene Cloning and Editing
Restriction enzyme example: EcoR1
Restriction site: GAATTC
CTTAAG
Restriction fragments: G cut A A T T C
C T T A A cut G
“Sticky Ends” – ss pieces of DNA fragments
whose unpaired bases can bind to
complementary ss regions of another DNA
fragment
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Gene Cloning and Editing
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Nucleic Acid Probes
Nucleic acid probe – a radioactive/fluorescent
labeled ss nucleic acid can be used to find a
specific gene or another nucleotide sequence
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Reverse Transcription
Eukaryotes have introns and exons while
prokaryotes do not have the machinery to remove
introns
Complementary DNA (cDNA) – a DNA molecule
made in vitro using a mRNA strand and reverse
transcriptase, corresponds to a gene without
introns
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CRISPR-Cas9 System
Cas9 protein – cuts double-stranded DNA at any
sequence directed by a molecule of RNA instead
of cutting a specific sequence like restriction
enzymes
Allows scientists to modify any genes without
altering any of the others
Cas9-guide RNA complex can target a specific
sequence such as a gene
Knock-out
Knock-in
Search and replace
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CRISPR-Cas9 System
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CRISPR-Cas9 System
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DNA Technology and Diseases
Treating Diseases
Humulin (produced by bacteria) – diabetes
HGH (produced by bacteria) – dwarfism
GBA (produced by carrot cells) – Gaucher’s disease
Diagnosing Diseases
Identifying inherited diseases before
symptoms/birth
Identifying precise strains responsible for
outbreaks
Preventing Diseases
Development of vaccines (harmless derivatives of
pathogens that stimulate an immune response
against the realCopyright
pathogen)
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DNA Technology and Diseases
Gene Therapy – a treatment for a disease/disorder in
which the patient’s defective gene is supplemented or
altered
Normal gene replaces defective gene then multiplied
throughout the body >>> stem cells
Is it ethical?
22 children treated for SCID (~25% had serious
side effects)
Hemophilia
Sickle-cell disease
Hereditary blindness
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DNA
Technology
and Diseases
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DNA Profiling
DNA Profiling – a procedure often used in the field of
forensics that analyzes DNA samples to determine
who they belong to
Most DNA is the same but there are polymorphic
regions that vary highly between people
Polymorphic regions can be selected, amplified, and
compared to match the regions – match = their DNA
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PCR
Polymerase Chain Reaction (PCR) – a technique used
to target a sequence of DNA and amplify it
PCR allows us to take a minute amount of a sample
and generate billions of copies (each cycle doubles
the DNA sequence amount)
Primers – short (~15-20), artificially created ssDNA
molecules that bind to the end of target sequences
for PCR
Key to high sensitivity and accurate results for PCR
Primers only bind to sequences pertaining to the
target so DNA polymerase (Taq polymerase) only
duplicates the desired target of DNA
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Gel Electrophoresis
Gel Electrophoresis – a technique used to separate
and analyze either DNA and proteins
Gel is made up of agarose
DNA is negative due to its phosphate group – When
placed in the well, DNA will travel toward the positive
end
DNA can be separated by lengths with shorter
molecules/segments traveling faster (further from
wells) while longer segments traveling slower (closer
to wells)
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Gel Electrophoresis
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STR Analysis
Short Tandem Repeats (STRs) – a series of short DNA
sequences that are repeated multiple times in a row
in a genome
CODIS 13 – repeats of four-nucleotide DNA
sequences at 13 sites through the human genome
Typically contains 3-50 repeats per site
# of repeats at each site is different for each
person
Random matching is 1 out of 10 billion
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STR Analysis
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Genomics
Genomics – the study of complete sets of genes and
their interactions (the study of whole genomes)
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Genomics
Human Genome Project – an international collaboration to map
and sequence the DNA of a human’s genome, started in 1990
and finished in 2004
Estimated genome size
3 billion nucleotide base pairs
21,000 genes
Estimated genome breakdown
1.5% - exons
5% - regulatory sequences
20% - introns
15% - unique noncoding DNA
15% - repetitive DNA unrelated to transposons
43.5% - repetitive DNA including transposons
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Genomics
Shotgun DNA Sequence Method – a method for
determining the DNA sequence of an entire genome
First genome = 13 years and $100 million
Today’s genome = a couple of hours and $600
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