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20 Φεβ 2013 (πριν από 4 χρόνια και 6 μήνες)

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Lecture # 3

Introduction & History of
Biotechnology

Recombinant Technology

Fundamentals of Biotechnology

By: Haji Akbar

M Phil


Biotechnology

is

most

briefly


“the

art

of

utilizing

living

organisms

and

their

products

for

the

production

of

food,

drink,

medicine

or

for

other

benefits

to

the

human

race,

or

other

animal

species

.






Humans

have

been

making

use

of

biotechnology

since

their

life

on

earth
.

e
.
g
.

Discovered

farming,



Animal

breeding



Cross
-
pollination

of

plants

and

cross
-
breeding

of

animals

(used

to

enhance

product

quality

and/or

meet

specific

requirements

or

standards)
.




Most

significant,

life
-
altering

discoveries
.


The

discovery

of

microorganisms

and

the

subsequent

burst

of

knowledge

related

to

the

causes

of

infectious

diseases,

antibiotics

and

immunizations
.


Current

scientific

methods

are

more

specific

than

historical

techniques,



as

scientists

now

directly

alter

genetic

material

using

techniques

known

as

recombinant

DNA

technology
.


Subjects Involved With Biotechnology


Multidisciplinary
-

involving

a

number

of

disciplines

that

are

coordinated

for

a

desired

outcome








Science


Life sciences


Physical sciences


Social sciences


Mathematics


Applied sciences


Computer applications


Engineering


Agriculture


Stages of Biotechnology Development


Ancient biotechnology
-

early history as
related to food and shelter; Includes
domestication



Classical biotechnology
-

built on ancient
biotechnology; Fermentation promoted
food production, and medicine



Modern biotechnology
-

manipulates
genetic information in organism; Genetic
engineering

Benefits of Biotechnology


Medicine


Human


Veterinary


Biopharming


Environment


Agriculture


Food products


Industry and manufacturing
etc.

The legends in Biotechnology


Anton van
Leeuwenhoek

(father of Microbiology)


Discovered cells


Gregor Johan Mendel


Discovered Genetics

(father of Genetics)


Walter Sutton






Discovered
Chromosomes


Thomas Hunt Morgan


Discovered how genes
are transmitted
through chromosomes


Sir Alexander Fleming


Discovered penicillin


Rosalind Elsie Franklin


Research led to the
discovery of the
double helix structure
of DNA


James Watson and
Francis Crick


Discovered DNA


Ernst Ruska


Invented the electron
microscope
(in 1933)
.


Mary
-
Claire King


Mapped human genes
for research of cancer
treatments


Ian Wilmut


Created the first true
clone, the Dorset ewe
Dolly

Recombinant DNA Technology


Recombinant

DNA

(rDNA)

technology

is

a

field

of

molecular

biology

in

which

scientists

"edit"

DNA

to

form

new

synthetic

molecules,

which

are

often

referred

to

as

"chimeras"
.




Recombinant

DNA

technology

works

by

taking

DNA

from

two

different

sources

and

combining

that

DNA

into

a

single

molecule
.




Recombinant

DNA

is

a

type

of

DNA

that

is

artificially

created

by

inserting

a

strand

or

more

of

DNA

into

a

different

set

of

DNA
.

often

referred

to

as

rDNA

for

short
.




Recombinant DNA technology

Recombinant

DNA

technology

is

one

of

the

recent

advances

in

biotechnology,

which

was

developed

by

two

scientists

named

Boyer

and

Cohen

in

1973
.



Basic principle of recombinant
DNA technology


The

DNA

is

inserted

into

another

DNA

molecule

called

‘vector



The

recombinant

vector

is

then

introduced

into

a

host

cell

where

it

replicates

itself

Steps involved recombinant DNA technology


1. DNA molecules are digested with
enzymes called
restriction
endonucleases

which reduces the size of the fragments


Renders them more manageable for cloning
purposes



Can be called
Restriction digestion


2. These products of digestion are inserted
into a DNA molecule called a vector




Enables desired fragment to be replicated in
cell culture to very high levels in a given cell
(copy #)



3. Introduction of recombinant DNA molecule
into an appropriate host cell


Transformation or transfection


Each cell receiving rDNA = CLONE


May have thousands of copies of rDNA
molecules/cell after DNA replication


As host cell divides, rDNA partitioned into
daughter cells



4. Population of cells of a given clone is expanded,
and therefore so is the
rDNA
.


Amplification


DNA can be extracted, purified and used for molecular
analyses


Investigate organization of genes


Structure/function


Activation


Processing


Gene product encoded by that
rDNA

can be characterized
or modified through mutational experiments




Applications of Recombinant DNA Technology


Large
-
scale

production

of

human

proteins

by

genetically

engineered

bacteria
.



Such

as

:

insulin,

Growth

hormone,

Interferons

and



Blood

clotting

factors

(VIII

&

IX)


Production of Human Insulin

1
)

Obtaining

the

human

insulin

gene


Human

insulin

gene

can

be

obtained

by

making

a

complementary

DNA

(
cDNA
)

copy

of

the

messenger

RNA

(mRNA)

for

human

insulin
.






2
)Joining

the

human

insulin

gene

into

a

plasmid

vector



The

bacterial

plasmids

and

the

cDNA

are

mixed

together
.

The

human

insulin

gene

(
cDNA
)

is

inserted

into

the

plasmid

through

complementary

base

pairing

at

sticky

ends
.



3
)Introducing

the

recombinant

DNA

plasmids

into

bacteria




The

bacteria

E
.
coli

is

used

as

the

host

cell
.

If

E
.

coli

and

the

recombinant

plasmids

are

mixed

together

in

a

test
-
tube
.




4
)Selecting

the

bacteria

which

have

taken

up

the

correct

piece

of

DNA




The

bacteria

are

spread

onto

nutrient

agar
.

The

agar

also

contains

substances

such

as

an

antibiotic

which

allows

growth

of

only

the

transformed

bacteria
.

(selective

media)




2.
Vaccine Development


3.
recombinant factor 8

and
recombinant
factor 9

made by genetic engineering are now
available for hemophilia A and B.


4.
Gene therapy for genetic diseases