1. Biology, 9th ed,Sylvia Mader
Chapter 16
Biotechnology and Genomics
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
DNA probe array
tagged DNA did
bind to probe
DNA probe
tagged DNA did not
bind to probe
tagged
DNA
testing subject's DNA

2. Biology, 9th ed,Sylvia Mader
Chapter 16
Outline
Biotechnology and Genomics
DNA Cloning
Recombinant DNA Technology
Restriction Enzyme
DNA Ligase
Polymerase Chain Reaction
Biotechnology Products
Transgenic Bacteria, Plants, and Animals
Gene Therapy
Genomics

3. Biology, 9th ed,Sylvia Mader
Chapter 16
DNA Cloning
Biotechnology and Genomics
Cloning is the production of identical copies of DNA
Members of a bacterial colony on a petri dish are clones because they all came from division of the same cell.
Identical twins are clones
Single embryo separate to become two.
Gene cloning is production of many identical copies of the same gene.
If the inserted gene is replicated and expressed, we can recover the cloned gene or protein product.
Cloned genes have many research purposes
Humans can be treated with gene therapy

4. Recombinant DNA Technology
Biology, 9th ed,Sylvia Mader
Chapter 16
Recombinant DNA Technology
Biotechnology and Genomics
Recombinant DNA (rDNA) contains DNA from two or more different sources
Requires:
A vector
introduces rDNA into host cell
Plasmids (small accessory rings of DNA from bacteria) are common vectors
Two enzymes are required to introduce foreign DNA into vector DNA
A restriction enzyme - cleaves DNA, and
A DNA ligase enzyme - seals DNA into an opening created by the restriction enzyme

5. Biology, 9th ed,Sylvia Mader
Chapter 16
Cloning a Human Gene
Biotechnology and Genomics
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
human DNA
plasmid
bacterium
human cell O f r o m a i
1. Restriction enzyme
cleaves DNA.
insulin gene
2. DNA ligase seals
human gene and
plasmid.
recombinant DNA
3. Host cell takes up
recombined plasmid.
4. Gene cloning occurs.
insulin

6. Biology, 9th ed,Sylvia Mader
Chapter 16
Restriction Enzyme
Biotechnology and Genomics
Cuts DNA at specific points.
Cleaves vector (plasmid) and foreign (human) DNA.
Cleaving DNA makes DNA fragments ending in short single-stranded segments with “sticky ends.”
The “sticky ends” allow insertion of foreign DNA into vector DNA.
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. A G A A T T C G C
DNA
duplex T C T T A A G C G
restriction
enzyme A A T T C G C
"sticky ends" G C G A G T C T T A A

7. Biology, 9th ed,Sylvia Mader
Chapter 16
DNA Ligase
Biotechnology and Genomics
Seals the foreign gene into the vector DNA
Treated cells (bacteria) take up plasmids
Bacteria and plasmids reproduce.
Many copies of the plasmid and many copies of the foreign gene.

8. DNA Cloning: Polymerase Chain Reaction (PCR)
Biology, 9th ed,Sylvia Mader
Chapter 16
DNA Cloning: Polymerase Chain Reaction (PCR)
Biotechnology and Genomics
Amplifies a targeted sequence of DNA
Create millions of copies of a single gene or a specific piece of DNA in a test tube
Requires:
DNA polymerase
Withstands the temperature necessary to separate double-stranded DNA.
A supply of nucleotides for the new, complementary strand

9. Biology, 9th ed,Sylvia Mader
Chapter 16
PCR
Biotechnology and Genomics
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
PCR
cycles
DNA
copies
first 1
DNA double strand
second 2
new strand
old strand
third 4
new
old
fourth 8
old
new
fifth 16
and so forth

10. Applications of PCR: Analyzing DNA Segments
Biology, 9th ed,Sylvia Mader
Chapter 16
Applications of PCR: Analyzing DNA Segments
Biotechnology and Genomics
DNA fingerprinting is the technique of using DNA fragment lengths
Treat DNA segment with restriction enzymes
A unique collection of different fragments is produced
Gel electrophoresis separates the fragments according to their charge/size
Produces distinctive banding pattern
Usually used to measure number of repeats of short sequences
Used in paternity suits, rape cases, corpse ID, etc.

11. DNA Fingerprinting & Paternity
Biology, 9th ed,Sylvia Mader
Chapter 16
DNA Fingerprinting & Paternity
Biotechnology and Genomics
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Mother
Child
Male 1
Male 2
few
Base repeat units
many a.
DNA Band patterns
Fluorescence units
Increasing size
b. Automated DNA fingerprinting

12. Biotechnology Products
Biology, 9th ed,Sylvia Mader
Chapter 16
Biotechnology Products
Biotechnology and Genomics
Genetically engineered organisms can produce biotechnology products.
Organisms that have had a foreign gene inserted into them are transgenic.

13. Biology, 9th ed,Sylvia Mader
Chapter 16
Transgenic Bacteria
Biotechnology and Genomics
Gene of interest is inserted into bacteria.
Bacteria are grown in large vats called bioreactors and product is harvested.
Products on the market include insulin, hepatitis B vaccine, t-PA, and human growth hormone.
Transgenic bacteria can produce chemical products.
Transgenic bacteria process minerals.
Metals Collection

14. Biology, 9th ed,Sylvia Mader
Chapter 16
Transgenic Bacteria
Biotechnology and Genomics
Transgenic bacteria promote plant health
Bacteria that colonize corn roots can be endowed with genes for insect toxin
Transgenic bacteria can degrade substances.
Oil-Eating Bacteria

15. Genetically Engineered Bacteria
Biology, 9th ed,Sylvia Mader
Chapter 16
Genetically Engineered Bacteria
Biotechnology and Genomics
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
(Both): Courtesy General Electric Research & Development

16. Biology, 9th ed,Sylvia Mader
Chapter 16
Transgenic Plants
Biotechnology and Genomics
Agricultural Crops
Foreign genes now give cotton, corn, and potato strains the ability to produce an insect toxin
Soybeans are now resistant to a common herbicide
Human Hormones
Plants are being engineered to produce human proteins including hormones, clotting factors, and antibodies in their seeds

17. Biology, 9th ed,Sylvia Mader
Chapter 16
Transgenic Animals
Biotechnology and Genomics
Vortex Mixing:
Many types of animal eggs have taken up the gene for bovine growth hormone (bGH)
The procedure has been used to produce larger fishes, cows, pigs, rabbits, and sheep
Gene Pharming:
Use of transgenic farm animals to produce pharmaceuticals
Genes coding for therapeutic & diagnostic proteins are incorporated into an animal’s DNA
The proteins appear in the animal’s milk
Plans are to produce drugs to treat
Cystic fibrosis
Cancer
Blood diseases, etc.

18. Biology, 9th ed,Sylvia Mader
Chapter 16
Transgenic Mammals
Biotechnology and Genomics
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
human gene
for growth
hormone
microinjection of human gene
donor of egg
development within
a host goat
human growth
hormone
Transgenic goat produces
human growth hormone.
milk a.
transgenic goat cells
with gene for human
growth hormone
microinjection of
transgenic gene into
enucleated donor eggs
enucleated eggs
donor of eggs
development
within
host goats
Cloned
transgenic
goats produce
human growth
hormone.
milk b.

19. Biology, 9th ed,Sylvia Mader
Chapter 16
Transgenic Animals
Biotechnology and Genomics
Researchers are using transgenic mice for various research projects.
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
one-celled
mouse embryos
with two X
chromosomes
no injection
inject SRY DNA
Embryo develops
into a female.
Embryo develops
into a male.
FEMALE
MALE

20. Biology, 9th ed,Sylvia Mader
Chapter 16
Gene Therapy
Biotechnology and Genomics
Gene therapy involves procedures to give patients healthy genes to make up for a faulty gene.
It also includes the use of genes to treat genetic disorders and various human illnesses.
There are ex vivo (outside body) and in vivo (inside body) methods of gene therapy.
Ex Vivo
Children with Severe Combined Immunodeficiency
Bone Marrow Stem Cells
In Vivo
Cystic Fibrosis
Nasal / Respiratory Spray

21. Biology, 9th ed,Sylvia Mader
Chapter 16
Gene Therapy
Biotechnology and Genomics
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Brain
(gene transfer by injection)*
• Huntington disease
• Alzheimer disease
• Parkinson disease
• brain tumors
Skin
(gene transfer by modified
blood cells)**
• skin cancer
Lungs
(gene transfer by aerosol spray)*
• cystic fibrosis
• hereditary emphysema
Liver
(gene transfer by modified
implants)**
• familial hypercholesterolemia
Blood
(gene transfer by bone
marrow transplant)*
• sickle-cell disease
Endothelium
(blood vessel lining)
(gene transfer by
implantation of
modified implants)**
• hemophilia
• diabetes mellitus
Muscle
(gene transfer by injection)*
• Duchenne muscular dystrophy
Bone marrow
(gene transfer by implantation of
modified stem cells)**
• SCID
• sickle-cell disease
* in vivo
** ex vivo

22. Biology, 9th ed,Sylvia Mader
Chapter 16
Genomics
Biotechnology and Genomics
Genomics is the study of genomes of humans and other organisms.
Sequencing the Bases
The Human Genome Project produced a working draft of all the base pairs in all chromosomes.
Took 13 years to sequence three billion base pairs along the length of chromosomes.

23. Biology, 9th ed,Sylvia Mader
Chapter 16
Human Genome Project
Biotechnology and Genomics
Genome - All the genetic information of an individual (or species)
Goals of Human Genome Project
Determine the base pair sequence
Construct a map showing sequence of genes on specific chromosomes

24. Biology, 9th ed,Sylvia Mader
Chapter 16
Human Genome Project
Biotechnology and Genomics
Humans have 20,500 genes
Most code for proteins
Much of the human genome was formerly described as “junk”
Does not specify the order of amino acids in a polypeptide
Recent observation suggest that between 74% and 93% of the genome is transcribed into RNA
Thus, vast junk DNA wasteland may be much more important than once thought

25. Eukaryotic Gene Structure
Biology, 9th ed,Sylvia Mader
Chapter 16
Eukaryotic Gene Structure
Biotechnology and Genomics
Historically, genes were defined as discrete units of heredity that corresponded to a locus on a chromosome.
Prokaryotes typically possess a single circular chromosome.
Eukaryotic chromosomes are much more complex.

26. Biology, 9th ed,Sylvia Mader
Chapter 16
Chromosomal DNA
Biotechnology and Genomics
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
intergenic sequences
Gene C
exon
exon
DNA
intron
Gene A
Gene B
exon
exon
pre-mRNA
intron
RNA
introns 5¢ 3¢ 5¢ 3¢ 3¢ 5¢
Gene A mRNA
Gene B mRNA
Gene C mRNA
© Cindy Charles/PhotoEdit

27. Eukaryotic Gene Structure
Biology, 9th ed,Sylvia Mader
Chapter 16
Eukaryotic Gene Structure
Biotechnology and Genomics
An Intergenic Sequences are DNA sequences that occur between genes
Repetitive DNA elements occur when the same sequence of two or more nucleotides are repeated many times along the length of one or more chromosomes.
Transposons are specific DNA sequences that have the remarkable ability to move within and between chromosomes.

28. Genomic Differences Between Species
Biology, 9th ed,Sylvia Mader
Chapter 16
Genomic Differences Between Species
Biotechnology and Genomics
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

29. Biology, 9th ed,Sylvia Mader
Chapter 16
HapMap Project
Biotechnology and Genomics
People inherit patterns of sequence differences, called haplotypes
If one haplotype of a person has an A rather than a G at a particular location in a chromosome, there are probably other particular base differences near the A
Genetic data from African, Asian, and European populations will be analyzed
A HapMap is a catalog common sequence differences that occur in a species
The goal of the project is to link haplotypes to risk for specific illnesses
May lead to new methods of preventing, diagnosing, and treating disease

30. Functional and Comparative Genomics
Biology, 9th ed,Sylvia Mader
Chapter 16
Functional and Comparative Genomics
Biotechnology and Genomics
Functional genomics aims to understand the role of genome in cells or organisms
DNA microarrays can monitor the expression of thousands of genes simultaneously and tell us:
What genes are turned on
Environmental conditions that turn on the gene
DNA microarrays contain microscopic amounts of known DNA fixed onto a small glass slide
mRNA bind through complementary base pairing
mRNA is produced by active cell.
Identify various mutations in the genome of an individual
This is called the person’s genetic profile.

31. DNA Microarray Technology
Biology, 9th ed,Sylvia Mader
Chapter 16
DNA Microarray Technology
Biotechnology and Genomics
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
DNA probe array
tagged DNA did
bind to probe
DNA probe
tagged DNA did not
bind to probe
tagged
DNA
testing subject's DNA

32. Biology, 9th ed,Sylvia Mader
Chapter 16
Genetic Profile
Biotechnology and Genomics
The complete genotype of an individual
This is the person’s genetic profile
A way of studying how genes work together to control the phenotype
Analyze the genetic profile of many individuals
Compare their profiles to their phenotypes

33. Biology, 9th ed,Sylvia Mader
Chapter 16
Proteomics
Biotechnology and Genomics
The study of the structure, function, and interaction of cellular proteins
At least 25,000 of our genes are translated into proteins
The sum total of these proteins is called the human proteome
Understanding protein function is essential to the development of better drugs
Correlate drug treatment to the particular genome
Increase efficiency and decrease side effects
Once the primary structure of these protein is known
It should be possible to predict their tertiary structure
Computer modeling of the tertiary of these proteins is an important part of proteomics

34. Biology, 9th ed,Sylvia Mader
Chapter 16
Bioinformatics
Biotechnology and Genomics
The application of computer technologies to the study of the genome
Genomics and proteomics produce raw data
These fields depend on computer analysis to find significant patterns in the data
Scientists hope to find relationships between genetic profiles and genetic disorders
New computational tools will be needed to accomplish these goals

35. Biology, 9th ed,Sylvia Mader
Chapter 16
Bioinformatics
Biotechnology and Genomics
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
intergenic sequences
Gene C
exon
exon
DNA
intron
Gene A
Gene B
exon
exon
pre-mRNA
intron
RNA
introns 5¢ 3¢ 5¢ 3¢ 3¢ 5¢
Gene A mRNA
Gene B mRNA
Gene C mRNA

36. Biology, 9th ed,Sylvia Mader
Chapter 16
Review
Biotechnology and Genomics
DNA Cloning
Recombinant DNA Technology
Restriction Enzyme
DNA Ligase
Polymerase Chain Reaction
Biotechnology Products
Gene Therapy
Genomics

37. Biology, 9th ed,Sylvia Mader
Chapter 16
Biotechnology and Genomics
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
DNA probe array
tagged DNA did
bind to probe
DNA probe
tagged DNA did not
bind to probe
tagged
DNA
testing subject's DNA