Biotechnology

aquaaniseΒιοτεχνολογία

6 Δεκ 2012 (πριν από 4 χρόνια και 7 μήνες)

333 εμφανίσεις


Greatest
Accomplishment has
been the sequencing of
the human genome.


Completed in 2003


Chromosome Maps


Applications


Pharm. Developments


Genetically altered mice


Gene therapy


Resist to herbicides and infection


Increased resistance to
env
.
Factors


Food with pharm.


Production of pharmaceuticals for treatment of
diseases.


E.g. Human Insulin, GH


Vaccines


Gene Therapy


Artifically replace disease causing gene with normal gene


Genetically modified organisms (GMO)


enabling plants to produce new proteins


Protect crops from insects
:
BT corn



corn produces a bacterial toxin that kills corn borer (caterpillar
pest of corn)


Extend growing season
:
fishberries



strawberries with an anti
-
freezing gene from flounder


Improve quality of food
:
golden rice



rice producing vitamin A

improves nutritional value


Bacteria review


one
-
celled prokaryotes


reproduce by binary fission


rapid growth


generation every ~20 minutes


10
8

(100 million) colony overnight!


incredibly diverse


Single circular chromosome


Plasmids


Transferable through transformation and conjugation


Restriction enzymes


Ligase


Plasmids/cloning


Gel electrophoresis


PCR


RFLPs


Used as
vectors

to insert new genes into bacteria

plasmid

cut DNA

gene from

other organism

glue DNA

recombinant

plasmid

vector

transformed

bacteria

Bacteria express
the new gene


radar

racecar

Madam I’m Adam

Able was I ere I saw Elba

a man, a plan, a canal, Panama

Was it a bar or a bat I saw?

go hang a salami I’m a lasagna hog

palindromes


Action of enzyme


cut DNA at specific sequences


restriction site


Many
different enzymes


named after organism they are found in


EcoR
I
,
Hind
III
,
BamH
I
,
Sma
I


Cut DNA at specific sites


leave “sticky ends”


GTAAC
G

AATTC
ACGCTT



CATTG
CTTAA G
TGCGAA

GTAAAC
GAATTC
ACGCTT

CATTTG
CTTAAG
TGCGAA

restriction enzyme cut site

restriction enzyme cut site

http://highered.mcgraw
-
hill.com/olc/dl/120078/bio37.swf



Cut other DNA with
same enzymes


leave “sticky ends”
on both


can glue DNA
together at



Since all living
organisms…


use the same DNA


use the same code
book


read their genes the
same way



Using restriction enzymes to build “customized”
plasmids


Antibiotic resistance genes in plasmid used as


Basically…DNA
Replication in a test
tube



http://www.sumanasinc.com/webcontent/
animations/content/pcr.html


http://www.youtube.com/watch?v=x5yPkx
CLads



The
are critical!


Primers have to
complement target
sequence


primers define section of
DNA to be cloned


Heat DNA to denature (unwind) it


90
°
C destroys DNA polymerase


have to add new enzyme every cycle


almost impractical!


Need enzyme that can

withstand 90
°
C…


from hot springs bacteria


Thermus

aquaticus


http://learn.genetics.utah.edu/content/labs/pcr
/



Uses:


Forensics


Evolutionary
Relationships


Paternity Testing


Medical Diagnostics



DNA is negatively
charged


DNA moves toward the
positive side


Smaller fragments
travel further



http://learn.genetics.utah.edu/content/labs/gel/


1

2

cut DNA with restriction enzymes

3

Stain DNA


ethidium

bromide
binds to DNA


fluoresces under UV
light



Comparing DNA samples from different organisms
to measure evolutionary relationships/similarity in
genetic makeup.



+

DNA



1

3

2

4

5

1

2

3

4

5

turtle

snake

rat

squirrel

fruitfly

chromosome with

disease
-
causing

allele 2

chromosome

with normal

allele 1



+

DNA



2.

RFLPs:
estriction
ragment
ength
olymorphism



G C T
T

G T A
A

C
G
G

C
C

T
C A T C A T C A T
T

C G C
C

G
G

C
C

T A C G C T
T


C G A
A

C A T
T

G
C
C

G
G

A
G T A
G

T AG T A
A G C G
G

C
C

G
G

A T G C G A
A


cut sites

cut sites

DNA





+

allele 1

G C T
T

G T A
A

C
G
G

C
C

T
C A T C A T C A T C A T C A T C A T
C
C

G
G

C
C

T A C G C T
T

C G A
A

C A T
T

G
C
C

G
G

A
G TA G T A G T A G T A G TA G TA
G
G

C
C

G
G

A T G C G A
A


allele 2

1

2

3


Evidence from murder
trial

+

DNA



child

Mom

F1

F2




Review:


Use of restriction enzymes


Transformation


PCR:

http://www.sumanasinc.com/webcontent/animations/content/pcr.html



Amplification of a DNA sample


Gel Electrophoresis
:
http://bcs.whfreeman.com/thelifewire/content/chp16/1601s.swf



Separation of DNA (proteins or RNA) based on size and charge


RFLP
:
http://highered.mcgraw
-
hill.com/olcweb/cgi/pluginpop.cgi?it=swf::535::535::/sites/dl/free/0072437316/120078/bio20.swf::Restriction%20Fragment%20Leng
th%
20P
olymorphisms



DNA Fingerprinting