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bio101
Citation preview
Sylvia S
. Mad
er
Copyright © The McGraw Hill Companies Inc. Permission required for reproduction or display
PowerPoint® Lecture Slides are prepared by Dr. Isaac Barjis, Biology Instructor
BIOLOGY10th Edition
Biotechnology and Genomics
Chapter14: pp. 249 - 263
1
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
DNA probe
DNA probe array
taggedDNA
tagged DNA didbind to probe
tagged DNA did notbind to probe
testing subject's DNA
2
Outline
DNA CloningRecombinant DNA Technology
Restriction EnzymeDNA Ligase
Polymerase Chain ReactionBiotechnology Products
Transgenic Bacteria, Plants, and Animals Gene TherapyGenomics
3
DNA Cloning
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
Animation
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5
Recombinant DNA Technology
Recombinant DNA (rDNA) contains DNA from two or more different sourcesRequires:
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
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7
Cloning a Human Gene
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
O from ai1. Restriction enzyme cleaves DNA.
2. DNA ligase seals human gene and plasmid.
3. Host cell takes uprecombined plasmid.
4. Gene cloning occurs.
bacterium
recombinant DNA
insulin
human cell
insulin gene
human DNA
plasmid
8
Restriction Enzyme
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.
DNAduplex
"sticky ends"
restrictionenzyme
A
T
A
T
A
T A
T
A
T
C
G
C
CG
G
C
G
A
T
A
T
A
T A
T
A
T
C
G
C
CG
G
C
G
Animation
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10
DNA Ligase
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.
Animation
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Animation
Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the “Normal” or “Slide Sorter” views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at http://get.adobe.com/flashplayer.
Animation
Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the “Normal” or “Slide Sorter” views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at http://get.adobe.com/flashplayer.
14
DNA Cloning: Polymerase Chain Reaction (PCR)
Amplifies a targeted sequence of DNACreate millions of copies of a single gene or a
specific piece of DNA in a test tube Requires:
DNA polymeraseWithstands the temperature necessary to separate
double-stranded DNA.A supply of nucleotides for the new,
complementary strand
15
PCR
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
PCRcycles
DNAcopies
first 1
second 2
third 4
old
old
old strand
new
new
new strand
DNA double strand
fourth 8
fifth 16
and so forth
Animation
16
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Animation
Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the “Normal” or “Slide Sorter” views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at http://get.adobe.com/flashplayer.
Animation
Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the “Normal” or “Slide Sorter” views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at http://get.adobe.com/flashplayer.
19
Applications of PCR: Analyzing DNA Segments
DNA fingerprinting is the technique of using DNA fragment lengthsTreat 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 patternUsually used to measure number of
repeats of short sequencesUsed in paternity suits, rape cases, corpse
ID, etc.
Animation
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21
DNA Fingerprinting & Paternity
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
Mother Child Male 1 Male 2
Bas
e re
pea
t u
nit
s
many
few
DNA Band patterns
Increasing size
b. Automated DNA fingerprinting
a.
Flu
ore
scen
ce u
nit
s
Animation
22
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23
Biotechnology Products
Genetically engineered organisms can produce biotechnology products.
Organisms that have had a foreign gene inserted into them are transgenic.
Animation
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25
Transgenic Bacteria
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
26
Transgenic Bacteria
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
27
Genetically Engineered Bacteria
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
(Both): Courtesy General Electric Research & Development
Animation
28
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29
Transgenic Plants
Agricultural CropsForeign genes now give cotton, corn, and
potato strains the ability to produce an insect toxin
Soybeans are now resistant to a common herbicide
Human HormonesPlants are being engineered to produce human
proteins including hormones, clotting factors, and antibodies in their seeds
Animation
30
Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the “Normal” or “Slide Sorter” views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at http://get.adobe.com/flashplayer.
31
Transgenic Animals
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.
32
Transgenic Mammals
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
human genefor growthhormone
human growthhormone
microinjection of human gene
development withina host goat
Transgenic goat produceshuman growth hormone.
a.
b.
transgenic goat cellswith gene for humangrowth hormone
microinjection oftransgenic gene intoenucleated donor eggs
donor of eggs
donor of egg
enucleated eggs
Clonedtransgenicgoats producehuman growthhormone.
milk
developmentwithinhost goats
milk
33
Transgenic Animals
Researchers are using transgenic mice for various research projects.
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
one-celledmouse embryos
with two Xchromosomes
Embryo developsinto a female.
FEMALE
Embryo developsinto a male.
MALE
no injection inject SRY DNA
34
Gene Therapy
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
35
Gene Therapy
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 modifiedblood cells)**• skin cancer
Lungs(gene transfer by aerosol spray)*• cystic fibrosis• hereditary emphysema
Liver(gene transfer by modifiedimplants)**• familial hypercholesterolemia
Blood(gene transfer by bonemarrow transplant)*• sickle-cell disease
Muscle(gene transfer by injection)*• Duchenne muscular dystrophy
Bone marrow(gene transfer by implantation ofmodified stem cells)**• SCID• sickle-cell disease
Endothelium(blood vessel lining)(gene transfer byimplantation ofmodified implants)**• hemophilia• diabetes mellitus
* in vivo
** ex vivo
36
Genomics
Genomics is the study of genomes of humans and other organisms.
Sequencing the BasesThe 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.
37
Human Genome Project
Genome - All the genetic information of an individual (or species)
Goals of Human Genome ProjectDetermine the base pair sequenceConstruct a map showing sequence of genes
on specific chromosomes
38
Human Genome Project
Humans have 20,500 genes Most code for proteinsMuch 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
39
Eukaryotic Gene Structure
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.
40
Chromosomal DNA
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
exon exon
intron
intron
exon exon
intergenic sequences
DNA
pre-mRNA
RNAintrons
Gene A Gene B
Gene C
Gene A mRNA Gene B mRNA Gene C mRNA
© Cindy Charles/PhotoEdit
5 3 5 3 3 5
41
Eukaryotic Gene Structure
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.
Animation
Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the “Normal” or “Slide Sorter” views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at http://get.adobe.com/flashplayer.
Animation
Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the “Normal” or “Slide Sorter” views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at http://get.adobe.com/flashplayer.
Animation
Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the “Normal” or “Slide Sorter” views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at http://get.adobe.com/flashplayer.
45
Genomic Differences Between Species
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
46
HapMap Project
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
47
Functional and Comparative 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.
48
DNA Microarray Technology
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
DNA probe
DNA probe array
taggedDNA
tagged DNA didbind to probe
tagged DNA did notbind to probe
testing subject's DNA
Animation
49
Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the “Normal” or “Slide Sorter” views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at http://get.adobe.com/flashplayer.
50
Genetic Profile
The complete genotype of an individualThis is the person’s genetic profileA way of studying how genes work together
to control the phenotypeAnalyze the genetic profile of many individualsCompare their profiles to their phenotypes
51
Proteomics
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
52
Bioinformatics
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
53
Bioinformatics
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
exon exon
intron
intron
exon exon
intergenic sequences
DNA
pre-mRNA
RNAintrons
Gene A Gene B
Gene C
Gene A mRNA Gene B mRNA Gene C mRNA
5 3 5 3 3 5
54
Review
DNA CloningRecombinant DNA Technology
Restriction EnzymeDNA Ligase
Polymerase Chain ReactionBiotechnology ProductsGene TherapyGenomics
Sylvia S
. Mad
er
Copyright © The McGraw Hill Companies Inc. Permission required for reproduction or display
PowerPoint® Lecture Slides are prepared by Dr. Isaac Barjis, Biology Instructor
BIOLOGY10th Edition
Biotechnology and Genomics
Chapter14: pp. 249 - 263
55
Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
DNA probe
DNA probe array
taggedDNA
tagged DNA didbind to probe
tagged DNA did notbind to probe
testing subject's DNA
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