14 Lecture Animation Ppt

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

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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

4

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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

Animation

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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

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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

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.

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

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.

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

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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

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.

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

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.

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

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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

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Genetically Engineered Bacteria

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

(Both): Courtesy General Electric Research & Development

Animation

28

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.

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

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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

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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

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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

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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.

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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

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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

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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

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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

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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