Biotechnology

Chapter 10

10.1 Impacts/Issues
Golden Rice or Frankenfood?

Genes from one species may be inserted into an

individual of another species or a gene may be
modified and reinserted into an individual of the
same species

Golden Rice
Rice plants with added genes make and store
beta carotene

Video: Golden rice or Frankenfood?

GMOs and Transgenic Organisms

Transgenic
An organism that has been genetically modified
with genes from a different species

Genetically modified organisms (GMOs)

Organism whose genome has been modified by
genetic engineering

10.2 Finding Needles in Haystacks

Gene research was limited until enzymes
produced by bacteria to cut viral DNA were
discovered
Restriction enzyme
Enzyme that cuts DNA at specific base sequences
Used in DNA cloning to cut DNA into pieces that
are inserted into cloning vectors

DNA Cloning
DNA cloning mass-produces DNA fragments for
research
DNA cloning
Set of procedures that uses living cells to make
many identical copies of a DNA fragment

Clone
A genetically identical copy of DNA, a cell, or an
organism

Cloning Vectors
Cloning vector
A DNA molecule that can accept foreign DNA,
resulting in a hybrid molecule that can be
transferred to a host cell, and get replicated in it

Plasmid
A small, circular DNA molecule in bacteria,
replicated independently of the chromosomes
A cloning vector

Recombinant DNA

Recombinant DNA molecules are introduced into

host cells such as bacteria, which copy the DNA
as they divide
Recombinant DNA
Contains genetic material from more than one
organism

Making Recombinant DNA

1. A restriction enzyme recognizes specific base
sequences in DNA from two different sources
2. Restriction enzymes cut DNA into fragments
with single-stranded tails (sticky ends)
3. DNA fragments from different sources are mixed
together; matching sticky ends base-pair

4. DNA ligase joins fragments, forming

recombinant DNA

Making Recombinant DNA

restriction
enzyme (cut)

1 A restriction enzyme
recognizes a specific base
sequence in DNA (green
boxes) from two sources.

mix

2 The enzyme cuts DNA

from both sources into
fragments that have
sticky ends.

DNA ligase
(paste)

3 The DNA fragments

from the two sources
are mixed together.
The matching sticky
ends base-pair with
each other.

4 DNA ligase joins

the fragments of
DNA where they
overlap. Molecules of
recombinant DNA
are the result.

Fig. 10-2, p. 181

restriction
enzyme (cut)

1 A restriction enzyme
recognizes a specific base
sequence in DNA (green
boxes) from two sources.

mix

2 The enzyme cuts DNA

from both sources into
fragments that have
sticky ends.

DNA ligase
(paste)

3 The DNA fragments

from the two sources
are mixed together.
The matching sticky
ends base-pair with
each other.

4 DNA ligase joins

the fragments of
DNA where they
overlap. Molecules of
recombinant DNA
are the result.

Stepped Art
Fig. 10-2, p. 181

Commercial Plasmid Cloning Vector

Kpn l
Sph l
Pst l
Bam Hl
Eco RI
Sal l

pDrive Cloning Vector

3.85 kb

Acc l
Xho l
Xba l
Bst XI

Sac l
Not l
Fig. 10-3, p. 181

cDNA Cloning
RNA cannot be cloned directly; reverse
transcriptase is used to copy single-stranded
RNA into cDNA for cloning
Reverse transcriptase
Viral enzyme that uses mRNA as a template to
make a strand of DNA

cDNA
DNA synthesized from an RNA template by the
enzyme reverse transcriptase

Making cDNA

mRNA

A The enzyme reverse

transcriptase transcribes
mRNA into DNA.
mRNA

cDNA
B DNA polymerase replicates
the DNA strand.
cDNA

cDNA
Eco RI recognition site
C The result is a double-stranded molecule
of DNA that can be cut and pasted into a
cloning vector.
Fig. 10-4, p. 182

mRNA

A The enzyme reverse

transcriptase transcribes
mRNA into DNA.
mRNA

cDNA
B DNA polymerase replicates
the DNA strand.
cDNA

cDNA
Eco RI recognition site
C The result is a double-stranded molecule
of DNA that can be cut and pasted into a
cloning vector.

Stepped Art
Fig. 10-4, p. 182

Libraries
A library is a collection of cells that host different
fragments of DNA, often representing an
organisms entire genome
Researchers make DNA libraries to isolate one
gene from the many other genes in a genome
Genome
An organisms complete set of genetic material

Nucleic Acid Hybridization

Probes are used to identify one clone that hosts
a DNA fragment of interest among many other
clones in a DNA library
Probe
Short fragment of DNA labeled with a tracer
Hybridizes with a specific nucleotide sequence

Nucleic acid hybridization

Base-pairing between DNA or RNA from different
sources

PCR
PCR quickly mass-produces copies of a
particular DNA fragment for study
Polymerase chain reaction (PCR)
Uses primers and heat-resistant DNA polymerase
to rapidly generate many copies of a DNA
fragment

Primer
Short, single-strand of DNA designed to hybridize
with a DNA fragment

Steps in PCR
1. Starting material is mixed with DNA polymerase,
nucleotides and primers
2. Mixture is heated and cooled in cycles
At high temperature, DNA unwinds
At low temperature, primers base-pair with
template DNA

3. Taq polymerase synthesizes complementary

DNA strands on templates

Two Rounds of PCR

1 DNA template (blue) is mixed with

primers (red), nucleotides, and heattolerant Taq DNA polymerase.

2 When the mixture is heated, the

double-stranded DNA separates
into single strands. zhen it is
cooled, some of the primers
base-pair with the template DNA.

3 Taq polymerase begins DNA

synthesis at the primers, and
complementary strands of DNA
form on the single-stranded
templates.

4 The mixture is heated again,

and the double-stranded DNA
separates into single strands.
When it is cooled, some of the
primers base-pair with the
template DNA.

5 Taq polymerase begins DNA

synthesis at the primers, and
complementary strands of DNA
form on the single-stranded
templates.
Fig. 10-5, p. 183

1 DNA template (blue) is mixed with

primers (red), nucleotides, and heattolerant Taq DNA polymerase.

22When
Whenthe
themixture
mixtureisisheated,
heated,the
the
double-stranded
double-strandedDNA
DNAseparates
separatesinto
into
single
singlestrands.
strands.When
Whenititisiscooled,
cooled,
some
someofofthe
theprimers
primersbase-pair
base-pairwith
withthe
the
template
templateDNA.
DNA.

44The
Themixture
mixtureisisheated
heatedagain,
again,
and
andthe
thedouble-stranded
double-strandedDNA
DNA
separates
separatesinto
intosingle
singlestrands.
strands.When
When
ititisiscooled,
cooled,some
someofofthe
theprimers
primers
base-pair
base-pairwith
withthe
thetemplate
templateDNA.
DNA.

3 Taq polymerase begins DNA

synthesis at the primers, and
complementary strands of DNA
form on the single-stranded
templates.

5 Taq polymerase begins DNA

synthesis at the primers, and
complementary strands of DNA
form on the single-stranded
templates.

Stepped Art
Fig. 10-5, p. 183

Animation: Polymerase chain reaction

(PCR)

Animation: Formation of recombinant

DNA

Animation: Use of a radioactive probe

Animation: Base-pairing of DNA

fragments

Animation: How to make cDNA

Animation: Restriction enzymes

Animation: F2 ratios interaction

10.3 Studying DNA

Short tandem repeats are multiple copies of a

short DNA sequence that follow one another
along a chromosome
The number and distribution of short tandem
repeats, unique in each individual, is revealed
by electrophoresis as a DNA fingerprint

DNA Fingerprinting
DNA fingerprinting is used in forensics, court
evidence, and other applications
DNA fingerprint
An individuals unique array of short tandem
repeats

Electrophoresis
Used to separate DNA fragments by size

DNA Fingerprinting: A Forensic Case

Suspect 2
Female Cells
Semen
Size Reference
Boyfriend
Control DNA
Control DNA
Size Reference

Control DNA
Size Reference
Victim
Suspect 1

Size Reference

Evidence from
Crime Scene

Fig. 10-6, p. 184

The Human Genome Project

Automated DNA sequencing and PCR enabled

scientists to sequence the more than 3 billion
bases of the human genome
Sequencing
Method of determining the order of nucleotides in
DNA

Sequencing a Fragment of DNA

The order of colors is the order of DNA bases (A,
T, G, C)

Genomics

Analysis of the human genome sequence is

yielding new information about human genes
and how they work
Genomics
The study of genomes (structural genomics,
comparative genomics)

Some Sequenced Genomes

Animation: Automated DNA sequencing

Animation: DNA fingerprinting

Video: ABC News: DNA mystery: Human

chimeras

Video: ABC News: Family ties: Paternity

testing

3D Animation: Gene sequencing

10.4 Genetic Engineering

Recombinant DNA technology and genome
analysis are the basis of genetic engineering
Genetic engineering is the directed alteration of
an individuals genome, resulting in a genetically
modified organism (GMO)
Genetic engineering
Process by which deliberate changes are
introduced into an individuals genome

Genetically Modified Microorganisms

A transgenic organism carries a gene from a
different species
Transgenic organisms are used in research,
medicine, and industry

Transgenic bacteria and yeast produce

medically valuable proteins

Designer Plants

Transgenic crop plants help farmers produce

food more efficiently

Plants with modified or foreign genes are now

common in farm crops

Using the Ti plasmid

to Make a Transgenic Plant

1 An A. tumefaciens bacterium
has been engineered to contain a
Ti plasmid. The plasmid carries a
foreign gene.
2 The bacterium infects a plant
cell and transfers the Ti plasmid
into it. The plasmid DNA becomes
integrated into one of the cells
chromosomes.
3 The plant cell divides, and its
descendants form an embryo.

4 The embryo develops into

a transgenic plant.

5 The transgenic plant

expresses the foreign
gene. This tobacco plant is
expressing a gene
from a firefly.
Fig. 10-8, p. 187

Animation: Gene transfer using a Ti

plasmid

Genetically Modified Crops

Bt gene confers insect resistance to corn

Biotech Barnyards
Transgenic animals produce human proteins

Animals that would be impossible to produce by

traditional breeding methods are being created
by genetic engineering
Transgenic animals are used in research,
medicine, and industry

Transgenic Animals

Knockout Cells and Organ Factories

Transgenic animals may one day provide a

source of organs and tissues for transplantation
into humans
Xenotransplantation
Transplant of an organ from one species to
another

Animation: Transferring genes into

plants

Video: ABC News: Cloned pooch

Video: ABC News: Mule clones

Video: ABC News: Glow-in-the-dark pigs

Video: ABC News: Cloned food approved

10.5 Genetically Modified Humans

Genes can be transferred into a persons cells to
correct a genetic defect or treat a disease
However, the outcome of altering a persons
genome remains unpredictable
Gene therapy
Transfer of a normal or modified gene into an
individual with the goal of treating a genetic
defect or disorder

Unpredictable Outcomes
There are more than 15,000 serious genetic
disorders gene therapy is the only real cure
In some cases, gene therapy works in other
cases it leads to death
Inserting a virus-injected gene into a
chromosome can disrupt normal function and
cause cancer
Severe allergic reaction to the viral vector can
cause death

One Successful Case of Gene Therapy

Rhys Evans, born with a severe immune
disorder (SCID-X1) received a normal gene and
no longer lives in isolation

Getting Perfect

Eugenics
Idea of deliberately improving the genetic
qualities of the human race

The potential benefits of genetically modifying

humans must be weighed against the potential
risks, including social implications

10.6 Impacts/Issues Revisited

Golden rice with beta carotene was ready for

distribution in 2005 but is still not available for
human consumption the biosafety experiments
required are too expensive for the public
humanitarian agency that developed it

Digging Into Data: Enhanced Spatial

Learning in Mice With Autism Mutation

Fig. 10-11a, p. 193

Fig. 10-11b, p. 193