1. MODERN
BIOTECHNOLOGY:
What
am
I
supposed
to
know…
MBB1
(1stAY2013-­‐2014)
MMP
Balolong

2. LECTURE
CONTENT:
PART
1
Ê DNA:
the
genetic
material
(ONE
GENE-­‐ONE
PROTEIN)
Ê Prokaryotic
system
Ê Eukaryotic
system
Ê INHERITANCE:
the
information
connection,
how
genes
are
passed
and
their
consequence
Ê Prokaryotic
system
Ê Eukaryotic
system

3. LECTURE
CONTENT:
PART
2
Ê MODERN
BIOTECHNOLOGY
Ê The
GMO
concept
Ê How
to
design/develop
a
GMO
Ê Tool/Methods:
Gene
Cloning,
Genetic
Engineering,
Recombinant
DNA
technology
Ê Equipment
and
Facilities
for
GMO
development
Ê Skills
and
Credentials

4. LECTURE
CONTENT:
PART
3
Ê MODERN
BIOTECHNOLOGY
:
advances
in
gene-­‐based
applications
Ê The
GMO
in
Health
and
Medicine
Ê The
GMO
in
Industry
Ê The
GMO
in
Food
and
Agriculture
Ê The
GMO
in
Environment

5. TARGET
ACTIVITIES
Ê WHO
KILLED
THE
VICTIM?
And
WHO’S
YOUR
PAPA?
(An
activity
on
Forensics)
Ê EXPOSURE
TRIP
(July
24,
2013)
Ê BIOTECH,
UPLB
Ê IRRI
Ê IPB
Ê PHIL
CARABAO
CENTER

6. www.crystalgraphics.com

7. THE
3
DOMAINS
OF
LIFE
bio1151b.nicerweb.net
gcps.desire2learn.com

8. BACTERIA
&
ARCHAEA
https://ib-­‐biology2010-­‐12.wikispaces.com

9. PROKARYOTES
VERSUS
EUKARYOTES

10. CHROMOSOME
&
DNA

11. DNA
&
THE
CENTRAL
DOGMA
www.labgrab.com

12. DNA:
THE
GENETIC
MATERIAL
Ê WHY
DNA?
Ê The
Griffith
Experiment
Ê Concept
of
a
transforming
principle
Ê The
Avery,
McLeod
and
McCarty
Experiment
Ê That
DNA
is
the
transforming
principle
Ê The
Hershey
and
Chase
Experiment
Ê Transduction
(via
phage)

13. GRIFFITH’S
EXPERIMENT:BACTERIAL
TRANSFORMATION

14. AVERY,
MCLEOD
&
MCCARTY:
DNA
AS
TRANSFORMING
PRINCIPLE
ap-­‐biology-­‐lab.wikispaces.com

15. HERSHEY
&
CHASE
EXPERIMENT:
TRANSDUCTION

16. SO
HOW
IS
DNA,
THE
GENETIC
MATERIAL,
USED
FOR
BIOTECHNOLOGY?

17. THE
SHIFT:
TRADITIONAL
TO
MODERN
Ê Nature’s
variation
Ê Color,
Size
Ê Traditional
breeding
can
give
us
additional
colors
and
sizes
Ê Modern
Biotechnology
can
give
you
MORE…
Ê Resistance
to
environmental
stress
Ê Taste
Ê Nutrition
kendralindell.wordpress.com

18. WHAT
WE
CAN
DO
WITH
BIOTECHNOLOGY…
nakedmaninthetree.wordpress.com
www.ent.iastate.edu
www.flickr.com
www.ishs.org

19. GROUP
WORK…
Ê One
GMO
per
group
Ê Research
on:
Ê What
problem
want
to
be
addressed
by
the
GMO
Ê How
was
it
developed
Ê What
were
the
problems
encountered
Ê How
were
they
resolved
(problems)?
Ê ISSUES….
Ê Open
Discussion
next
meeting…

20. LAST
MEETING…
DNA
and
its
importance
in
BIOTECHNOLOGY
1.
Golden
Rice;
2.
BT
Corn;
3.
Edible
Vaccine
in
Potato;
4.
GMO
Soya;
5.
Recombinant
Hepatitis
B
vaccine

21. RECALL…IN
MODERN
BIOTECHNOLOGY…
WE
CAN
CONTROL
THE
VARIATION
Ê NOTE:
Ê Allelic
differences
at
genes
control
a
specific
trait
Ê GENE:
a
piece
of
DNA
that
controls
the
expression
of
a
trait
Ê ALLELE:
alterative
form
of
the
gene
(they
can
segregate
to
form
the
variation
naturally)

22. DNA
(gene)
RNA
Protein
Trait
(or
phenotype)
Transcription
Translation
Plant
height
Seed
shape
Traditional
Breeding
Biotechnology
a. Gene
manipulation
b. Gene
Introduction

23. Ê GMO
or
Genetically-­‐Modified
Organisms
Ê Organisms
with
ARTIFICIALLY-­‐ALTERED
DNA
Ê Foreign
gene
is
inserted
from
another
species:
(enables
GMO
to
express
the
trait
coded
by
the
new
gene)
=
TRANSGENICS
Ê An
existing
gene
is
altered:
to
make
it
express
at
a
higher
level
or
in
a
different
way
=
FOR
GENE
THERAPY
Ê Gene
is
deleted
or
deactivated:
to
prevent
the
expression
of
a
trait
(e.g.
delayed
ripening)

25. THE
GOLDEN
RICE
Ê WHY:
to
address
Vitamin
A
deficiency
problem
Ê Linked
to
blindness
Ê Influences
Diarrhea
and
Measles
Ê >100M
children
affected
Ê Vitamin
Delivery
as
PILLS
=
most
countries
no
access
Ê Alternative
=
vitamin
into
CROPS
www.goldenrice.org

26. β-­‐Carotene
Pathway
in
Plants
IPP
Geranylgeranyl
diphosphate
Phytoene
Lycopene
β
-­‐carotene
(vitamin
A
precursor)
Phytoene
synthase
Phytoene
desaturase
Lycopene-­‐beta-­‐cyclase
ξ-­‐carotene
desaturase
Problem:
Rice
lacks
these
enzymes
Normal
Vitamin
A
“Deficient”
Rice
(Isopentenyl
diphosphate)

27. The
Golden
Rice
Solution
IPP
Geranylgeranyl
diphosphate
Phytoene
Lycopene
β
-­‐carotene
(vitamin
A
precursor)
Phytoene
synthase
Phytoene
desaturase
Lycopene-­‐beta-­‐cyclase
ξ-­‐carotene
desaturase
Daffodil
gene
Single
bacterial
gene;
performs
both
functions
Daffodil
gene
β-­‐Carotene
Pathway
Genes
Added
Vitamin
A
Pathway
is
complete
and
functional
Golden
Rice

28. THOUGH
METABOLIC
PATHWAYS
ARE
COMPLEX……THEY
ARE
INTERRELATED
Understanding
pathways
is
critical
to
developing
new
products

29. Modifying
Pathway
Components
Can
Produce
New
Products
Modified
Lipids
=
New
Industrial
Oils
Turn
On
Vitamin
Genes
=
Relieve
Deficiency
Increase
amino
acids
=
Improved
Nutrition

30. DEVELOPING
OUR
GMOs:
Tools
that
you
can
USE
Ê Genetic
Manipulation
or
Genetic
Engineering
Ê GOAL:
introduce,
alter,
delete
Ê Cloning
and
Cell
Culture
Ê What
to
manipulate:
Genes,
Cells,
Tissue
Ê GENOMICS,
PROTEOMICS,
METABOLOMICS

32. www.earthtimes.org

33. METHODS
IN
GENETIC
ENGINEERING
Ê GENE
ISOLATION
Ê TRANSFORMATION
Ê SELECTION
Ê REGENERATION
Ê CONFIRMATION
OF
EXPRESSION

34. EXAMPLE:
INSULIN
BY
BIOTECHNOLOGY
muirbiology.wordpress.com

35. EXAMPLE:
BT
CORN
www.scq.ubc.ca
www.scq.ubc.ca

36. GENE
ISOLATION
globalgenes.org
www.bluesci.org
www.cvillepodcast.com
hms.harvard.edu

37. THE
PRINCIPLE
TO
ISOLATE
THE
GENE:
Complementary
Genetics
1.
Protein
sequence
is
related
to
gene
sequence
NH3
+-Met-Asp-Gly--------------Trp-Ser-Lys-COO-
ATG GAT-GCT TGG-AGT-AAA
C C C G
A TCT
G C
A
G
2.
The
genetic
code
information
is
used
to
design
PCR
primers
Forward
primer:
5ʼ’-­‐ATGGAT/CGCN-­‐3ʼ’
Reverse
primer:
5ʼ’-­‐T/CTTNC/GT/ACCA-­‐3ʼ’
Notes:
T/C
=
a
mixture
of
T
and
C
at
this
position;
N
=
a
mixture
of
all
four
nucleotides
Reverse
primer
is
the
reverse
complement
of
the
gene
sequence

38. THE
GENETIC
CODE
plato.stanford.edu

39. 3.
Use
PCR
to
amplify
gene
fragment
a.
template
DNA
is
melted
(94C)
3ʼ’
5ʼ’
5ʼ’
3ʼ’
3ʼ’
5ʼ’
5ʼ’
3ʼ’
b.
primers
anneal
to
complementary
site
in
melted
DNA
(55C)
3ʼ’
5ʼ’
5ʼ’
3ʼ’
3ʼ’
5ʼ’
5ʼ’
3ʼ’
c.
two
copies
of
the
template
DNA
made
(72C)

40. Denaturation:
DNA
melts
Annealing:
Primers
bind
Extension:
DNA
is
replicated
PCR
Animation

41. Human
clone
library
Clones
transferred
to
filter
PCR
fragment
probe
added
to
filter
Hot-­‐spots
are
human
gene
of
interest
4.
Gene
fragment
used
to
screen
library

42. HOW
WERE
YOUR
GENES
ISOLATED?
Ê Golden
Rice
Ê BT
Corn
Ê Edible
Vaccine
in
Potato
Ê GMO
Soya
Ê Recombinant
Hepatitis
B
vaccine

43. www.designingdigitally.com