biotechnology

books
Crommelin, J.A., and Sindelar, R.D., 1997, Pharmaceutical
Biotechnology,
Harwood
Academis
Publishers,
Amsterdam.
Glick, R.B., and Pasternack, J.J., 1998, Molecular
Biotechnology : Principle and Application of recombinant
DNA, ASM Press, Washington D.C.
Prave, P., Faust, U., Sittig, W., Sukatsch, D.A., 1989, Basic
Biotecnology A Strudents Guide, VCH Publishers,
Germany.

introduction

what is biotechnology?

using scientific methods with organisms to produce

new products or new forms of organisms

any technique that uses living organisms or

substances from those organisms to make or modify a
product, to improve plants or animals, or to develop
micro-organisms for specific uses

OECD (the Organisation of Economic Co-operation and

Development) defines biotechnology as " the
application of scientific and engineering principles to
the processing of materials by biological agents ".

what is biotechnology?

manipulation of genes is called genetic engineering or

recombinant DNA technology

genetic engineering involves taking one or more genes

from a location in one organism and either

Transferring them to another organism

Putting them back into the original organism in different
combinations

what is biotechnology?

cell and molecular biology

microbiology
genetics
anatomy and physiology
biochemistry
engineering
computer Science

applications

virus-resistant crop plants and livestock

diagnostics for detecting genetic diseases
acquired diseases
therapies that use genes to cure diseases
recombinant vaccines to prevent disease
biotechnology can also aid the environment

and

goals of biotechnology

To understand more about

inheritance and gene expression

To provide better understanding & treatment of

various diseases, particularly genetic disorders

To generate economic benefits, including improved

plants and animals for agriculture and efficient
production of valuable biological molecules

the

processes

Example: Vitamin A fortified engineered rice

of

biotechnology terms

Genome or Genomics

Transcriptome

DNA
RNA or portion of genome transcribed

Proteome or Proteomics

Proteins

types of biotechnology

Recombinant, R protein, R DNA

Genetically Modified Organism (GMO)
Antibody (monoclonal antibody)
Transgenic
Gene therapy, Immunotherapy
Risks and advantages of biotech

history

biotechnology development

Ancient biotechnology- early history as related to food

and shelter; Includes domestication

Classical
biotechnologybuilt
on
ancient
biotechnology;
Fermentation
promoted
food
production, and medicine

Modern
biotechnologymanipulates
information in organism; Genetic engineering

genetic

biotechnology

any technique that uses living organisms or

substances to make or modify a product, to improve
plants, animals, or microorganisms for specific uses

evolving corn

ancient biotech
History of Domestication and Agriculture
Paleolithic peoples began to settle and develop
agrarian societies about 10,000 years ago
Early farmers in the Near East cultivated wheat,
barley, and possibly rye
7,000 years ago, pastoralists roamed the Sahara
region of Africa with sheep, goats, cattle, and also
hunted and used grinding stones in food preparation
Early farmers arrived in Egypt 6,000 years ago with
cattle, sheep, goats, and crops such as barley,
emmer, and chick-pea

ancient biotech

Not sure why peoples began to settle down and

become sedentary

May be in response to population increases and the

increasing demand for food
Shifts in climate
The dwindling of the herds of migratory animals
Early Farmers could control their environment when
previous peoples could not

People collected the seeds of wild plants for

cultivation and domesticated some species of wild
animals living around them, performing selective
breeding

stone sheep, 2900 BC

ancient plant germplasm

The ancient Egyptians saved seeds and tubers, thus

saved genetic stocks for future seasons

Nikolai Vavilov, a plant geneticist, came up with first

real plan for crop genetic resource management

National Seed Storage Laboratory in Fort Collins,

Colorado is a center for germplasm storage in the U.S.

Agricultural expansion and the use of herbicides has

put germplasm in danger and led to a global effort to
salvage germplasm for gene banks

fermented food, 1500 BC

Yeast - fruit juice

wine
Brewing beer - CO2
Baking bread, alcohol
Egyptians used yeast in 1500 B.C.
1915-1920 Bakers Yeast

fermented food, 1500 BC

fermentation

Fermentation:
microbial
process
in
which
enzymatically controlled transformations of organic
compounds occur
Fermentation has been practiced for years and has
resulted in foods such as bread, wine, and beer
9000 B.C. - Drawing of cow being milked Yogurt - 4000
B.C. Chinese Cheese curd from milk - 5000-9000 years
ago

classical biotech

Describes the development that fermentation has taken place

from ancient times to the present

Top fermentation - developed first, yeast rise to top

1833 - Bottom fermentation - yeast remain on bottom

1886 Brewing equipment made by E.C. Hansen and still used

today

World War I fermentation of organic solvents for explosives

(glycerol)

World War II bioreactor or fermenter:

Antibiotics

Cholesterol Steroids

Amino acids

classical biotech

large quantities of vinegar

are
produced
by
Acetobacter on a substrate
of wood chips
fermented fruit juice is
introduced at the top of
the
column
and
the
column is oxygenated from
the bottom

classical biotech advances

In the 1950s, cholesterol was converted to cortisol

and sex hormones by reactions such as microbial
hydroxylation (addition of -OH group)

By the mid-1950s, amino acids and other primary

metabolites (needed for cell growth) were produced,
as well as enzymes and vitamins

By the 1960s, microbes were being used as sources of

protein and other molecules called secondary
metabolites (not needed for cell growth)

classical biotech advances

Today many things are produced:

Pharmaceutical compounds such as antibiotics

Amino Acids
Many chemicals, hormones, and pigments
Enzymes with a large variety of uses
Biomass for commercial and animal consumption (such
as single-cell protein)

amino acids and their uses

old biotech meets new

Fermentation and genetic engineering have been used

in food production since the 1980s
Genetically engineered organisms are cultured in
fermenters and are modified to produce large
quantities of desirable enzymes, which are extracted
and purified
Enzymes are used in the production of milk, cheese,
beer, wine, candy, vitamins, and mineral supplements
Genetic engineering has been used to increase the
amount and purity of enzymes, to improved an
enzymes function, and to provide a more costefficient method to produce enzymes.

Chymosin, used in cheese production, was one of the first

produced

foundations of modern biotech

1590 - Zacharias Janssen - First two

lens microscope (30x)

1665 - Robert Hooke - Cork Cellulae

(Small Chambers)

Anthony van Leeuwenhoek (200x)

1676 - animalcules (in pond water)
1684 - protozoa/fungi

microscopy

van Leeuwenhoeks microscope (200x)

van Leeuwenhoeks drawing of yeast

published in 1684

foundations of modern biotechnology

1838, Matthias Schleiden, determined that all plant

tissue was composed of cells and that each plant
arose from a single cell

1839, Theodor Schwann, came to

determination as Schleiden, for animals

1858, Rudolf Virchow, concluded that all cells arise

from cells and the cell is the basic unit of life

Before cell theory the main belief was vitalism: whole

organism, not individual parts, posses life

By the early 1880s, microscopes, tissue preservation

technology, and stains allowed scientists to better
understand cell structure and function

similar

transforming principle

1928 - Fred Griffith performed

experiments using Streptococcus
pneumonia
Two strains:
Smooth (S) - Virulent (gel coat)
Rough (R) - Less Virulent
Injected R and heat-killed S - mice
died and contained S bacteria
Unsure of what changed R to S,
which he called the Transforming
principle

transforming principle

1952 Alfred Hershey and Martha Chase

Used T2 bacteriophage, a virus that infects bacteria

Radiolabeled the bacteriophage with S35 (Protein) and
P32 (DNA)
Bacterial cells were infected and put in a blender to
remove phage particles
Analysis showed labeled DNA inside the bacteria and
was the genetic material

1952 Alfred Hershey and Martha Chase

1953 watson and crick

Determined the structure of

DNA
Rosalind Franklin and Maurice
Wilkins
provided
X-ray
diffraction data
Erwin Chargaff determined
the ratios of nitrogen bases
in DNA
DNA replication model - 1953
DNA bases made up of purine
and pyrimidine
Nobel Prize - 1962

first recombinant DNA experiments

1971 scientists manipulated

DNA and placed them into
bacteria

1972 scientists joined two DNA

molecules
from
different
sources
using
the
endonuclease EcoRI (to cut)
and DNA ligase (to reseal)

first recombinant DNA experiments

Herbert Boyer later went to Cold Spring Harbor

Laboratories and discovered a new technique
called gel electrophoresis to separate DNA
fragments

A current is applied so that the negative

charged DNA molecules migrate towards
the positive electrode and is separated by
fragment size

first recombinant DNA experiments

biotech revolution: cracking the code

1961, Nirenberg and Mattei made the

first attempt to break the genetic
code, using synthetic messenger
RNA (mRNA)
Nirenberg and Leder developed a
binding assay that allowed them to
determine
which
triplet
codons
specified which amino acids by using
RNA sequences of specific codons

first DNA cloning

Boyer, Helling Cohen, and Chang

joined DNA fragments in a vector,
and transformed an E. coli cell

Cohen and Chang found they could

place bacterial DNA into an unrelated
bacterial species

In 1980 Boyer and Cohen received a

patent for the basic methods of DNA
cloning and transformation

public reaction

Recombinant DNA technology sparked debates more

than 30 years ago among scientists, ethicists, the
media, lawyers, and others
In the 1980s it was concluded that the technology had
not caused any disasters and does not pose a threat to
human health or the environment

public reaction
However, concerns have focused on both applications
and ethical implications:
Gene therapy experiments have raised the question
of eugenics (artificial human selection) as well as
testing for diseases currently without a cure
Animal clones have been developed, and fears have
been expressed that this may lead to human cloning
In agriculture, there is concern about gene
containment and the creation of super weeds
(herbicide and/or pesticide resistant weeds)
Today, fears have focused on genetically engineered
foods in the marketplace and has resulted in the
rapid growth of the organic food industry

public reaction

progress continues

Many genetically modified disease, pest, and

herbicide-resistant plants are awaiting approval for
commercialization
Genes involved in disease are being identified
New medical treatments are being developed
Molecular pharming, where plants are being used to
produce pharmaceuticals (biopharmaceuticals), is
being developed

biotechnology

biotechnology

Biotechnology helps to meet our basic needs.

Food, clothing, shelter, health and safety

Improvements by using science

Science helps in production plants, animals and other
organisms

Also used in maintaining a good environment that

promotes our well being

biotechnology

Using scientific processes to get new organisms or

new products from organisms.

biotechnology

Large area
Includes many approaches and methods in science
and technology

official definition

Any technique that uses living organisms or

substances from those organisms to make or modify a
product, to improve plants or animals.
Or to develop microorganisms for specific uses.

agricultural view

All of the applied science based operations

producing food, fiber, shelter, and related products

Milk production
New horticultural and ornamental plants
Wildlife,
aquaculture,
natural
resources
environmental management

in

and

organismic biotech

Working with complete, intact organisms or their cells

Organisms are not genetically changed with artificial
means

Help the organism live better or be more productive

Goal improve organisms and the conditions in which
they grow

organismic biotech

Study and use natural genetic variations

Cloning is an example of organismic biotech

cloning

Process of producing a new organism from cells or

tissues of existing organism.
1997 cloned sheep Dolly in Edinburgh Scotland

molecular biotech

Changing the genetic make-up of an organism

Altering the structure and parts of cells
Complex!

molecular biotech

Uses genetic engineering, molecular mapping and

similar processes

genetic engineering

Changing the genetic information in a cell

Specific trait of one organism may be isolated,cut, and
moved into the cell of another organism

transgenic

Results of Gen. Eng. Are said to be transgenic

Genetic material in an organism has been altered

model organism

sizes

viruses

proteins involved in DNA, RNA,

protein synthesis
gene regulation
cancer and control of cell
proliferation
transport of proteins and
organelles inside cells
infection and immunity
possible
gene
therapy
approaches

bacteria

proteins involved in DNA, RNA,

protein synthesis, metabolism
gene regulation
targets for new antibiotics
cell cycle
signaling

yeast
Saccharomyces cerevisiae
control of cell cycle and cell
division
protein
secretion
and
membrane biogenesis
function of the cytoskeleton
cell differentiation
aging
gene
regulation
and
chromosome structure

roundworm
Caenorhabditis elegans
development of the body plane
cell lineage
formation and function of the
nervous system
control of programmed cell
death
cell proliferation and cancer
genes
aging
behaviour
gene
regulation
and
chromosome structure

fruit fly
Drosophila melanogaster
development of the body plane
generation of differentiated cell
lineages
formation
of
the
nervous
system, heart and musculature
programmed cell death
genetic control of behaviour
cancer genes and control of cell
proliferation
control of cell polarisation
effect of drugs, alcohol and
pesticides

fruit fly

fruit fly
EMBRYO
Body
segments

LARVA

Gene expression

ADULT FLY
Head end
Tail end

zebrafish

development of vertebrate
body tissue
formation and function of
brain and nervous system
birth defect
cancer

zebrafish

mice

development of body tissues

function
of
mammalian
immune system
formation and function of
brain and nervous system
models of cancer and other
human diseases
gene
regulation
and
inheritance
infectious disease

homeotic genes
Fly chromosomes

Mouse chromosomes

The order of
homeotic genes
is the same
The gene order
corresponds to
analogous body
regions
Fruit fly embryo (10 hours)

Adult fruit fly

Mouse embryo (12 days)

Adult mouse

mouse with human ear

plants

development and patterning

of tissues
genetics of cell biology
agricultural applications
physiology
gene regulation
immunity
infectious disease

genome specification
Organism

Type

Chromo Gene # (bp)

some #

Genome Size

Hepatitus B

virus

3215

E. coli

bacterium

4,394

4,639,221

S.cerevisiae

yeast

16

6,183

12,000,000

D. melanogaster

fruit fly

14,000

140,000,000

C. elegans

nematode

19,000

90,000,000

A. thaliana

plant

25,000

125,000,000

M.musculus

mouse

20

35,000

3,000,000,000

H. sapiens

human

23

35,000

3,000,000,000

genome specification

production

products of biotech

products of biotech

applications
Agriculture
Plant breeding to improve resistance to pests,
diseases, drought and salt conditions
Mass propagation of plant clones
Bioinsecticide
development modification of plants to improve
nutritional and processing characteristics
Chemical Industry
Production of bulk chemicals and solvents such as
ethanol, citric acid, acetone and butanol
Synthesis of fine specialty chemicals such as enzymes,
amino acids, alkaloids and antibiotics

applications
Medicine
Development of novel therapeutic molecules
medical treatments
Diagnostics
Drug delivery systems
Tissue engineering of replacement organs
Gene therapy

for

applications
Food Industry
Production of bakers' yeast, cheese, yogurt and
fermented foods such as vinegar and soy sauce
Brewing and wine making
Production of flavors and coloring agents
Veterinary Practice
Vaccine production
Fertility control
Livestock breeding

applications
Environment
Biological recovery of heavy metals from mine tailings
and other industrial sources
Bioremediation of soil and water polluted with toxic
chemicals
Sewage and other organic waste treatment

future of medicine

smart drugs for cancer and autoimmune diseases

(arthritis, psoriasis, diabetes)
gene-based diagnostics and therapies
pharmaco-genomics and personalised medicine
stem cells and regenerative medicine
health and longevity

the promise of biotech

DNA

protein

drugs are so complex they can only be synthesized in a living system

tools

recombination and crossover

recombination and crossover

recombination and crossover

If no exchange of genes (i.e.
phenotypic marker) occurs,
recombination event can not
be detected

recombination and crossover

cloning DNA
Insert the DNA into plasmids
Gene of interest is inserted into small DNA molecules known as
plasmids, which are self-replicating, extrachromosomal genetic
elements originally isolated from the bacterium, Escherichia coli.
The circular plasmid DNA is opened using the same endonuclease
that was used to cleave the genomic DNA.

Join the ends of DNA with the enzyme, DNA ligase.

The inserted DNA is joined to the plasmid DNA using another
enzyme, DNA ligase, to give a recombinant DNA molecule. The
new plasmid vector contains the original genetic information for
replication of the plasmid in a host cell plus the inserted DNA.

cloning DNA
Introduce the new vector into host
The new vector is inserted back into a host where many copies of
the genetic sequence are made as the cell grows and divide with
the replicating vector inside.

Isolate the newly-synthesized DNA or the protein coded for by the

inserted gene.
The host may even transcribe and translate the gene and
obligingly produce product of the inserted gene. Alternatively,
many copies of the DNA gene itself may be isolated for
sequencing the nucleic acid or for other biochemical studies.

cloning DNA

cloning DNA

cloning DNA

cloning DNA

electrophoresis

electrophoresis

electrophoresis

IfDNAistoolargeforconventionalelectrophoresis.
electrophoresis

bioprocess control

control

so where are the computers?

convergence of biotech and information technology

automated sequencing (Celera)

gene chips and microarrays
high throughput screening
data visualisation and data mining
web-based clinical trials and FDA submission
in silico simulations of biological systems

control

control

molecular modelling
simulation
bioinformatics
monitoring
expert systems

synthetic biology

synthetic biology

Creating lifelike characteristics through the use of

chemicals
Based on creating structures similar to those found in
living organisms

synthetic biology

Is important because it brings science closer to

creating life in the lab
Cells and tissues may be developed to treat human
injury and disease

synthetic biology

Synthetic biology hopes to bring engineering practices

common in other engineering disciplines to the field of
molecular genetics and thus create a novel nanoscale
computational substrate

Advantages

Tightly integrated biological inputs and outputs

Easily grow thousands of computational engines
Natural use of directed evolution

Disadvantages

Speed is on the order of millihertz (tens of seconds)

Modest computational capability of each engine

at MIT, Knights group

synthetic biology applications

Autonomous biochemical sensors

Biomaterial manufacturing
Programmed therapeutics
Smart agriculture
Engineered experimental systems for biologists

biotechnology?

the end