Recombinant DNA

crunchkingofprussiaBiotechnology

Dec 6, 2012 (4 years and 11 months ago)

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Entry Task 1/26


How many years do you think it will be
before the Genetics depicted in
GATTACA can be used in real life?



Turn in GATTACA Questions

Genetics TEST


Out of 40 points...



This WILL be on the semester Final…
…so lets Review!

DNA

Technology


Write down the
Yellow

Stuff.

DNA Technology


Scientists can manipulate DNA in
some important ways


DNA technologies have helped
advance all types of scientific
research and medical advancement


DNA Techonogy


Biotechnology: Using biology for industry or
manufacturing


Pharmaceuticals & Medicine, Farming &
Agriculture, Lots of other stuff…


DNA technologies


DNA Sequencing


PCR


Polymerase Chain Reaction


Genetic Engineering
-

DNA Recombination and
Transformation

Manipulating DNA


We can “Cut” DNA


Separate DNA molecules


“Read” DNA

Manipulating DNA


Restriction Enzymes “Cut” DNA


Recognize a specific sequence and cuts the
Molecule in half at that point.


Gel Electrophoresis is a method for
Separating DNA


Electricity pushes DNA through a “Gel” that
separates it based on Length.


Long strands of DNA move slowly through
Gel, small strands move quickly through
holes in Gel.


Gel Electrophoresis

Manipulating DNA


PCR makes new DNA molecules in High
quantity.


Add enzymes & Nucleotides to DNA and
new DNA molecules will be made


When Sequencing we add Labeled
nucleotides that STOP the formation of DNA


DNA Sequencing makes many strands of
varying length and we can identify the base at
the END of each strand

Human Genome Project…


Sequencing the Human Genome is not
as easy as putting DNA in a tube, and
sequencing it.


Human Genome is 3 BILLION base pairs
long
. (that’s 786 megabytes of data)


Sequencing is only affective for 300
-
1000 bases at a time


Short segments had to be sequenced
then overlapped to fit together
.

Human Genome Project…


Began in 1990…


Human Genome sequence was
completed in 2003


Now the focus is on determining which
portions code for which traits.

Some Vocabulary


Bioinformatics
: Analyzing Biological Data
(usually DNA or Protein sequences)


Recombinant DNA
: mixing DNA from different
organisms
(we do this a lot in research and will
explain more next week)


Plasmid
: a loop of DNA found in Prokaryotes
(bacteria)


Animal DNA is linear and makes chromosomes


Selective Breeding
: Breeding for particular
traits
(Yes this is biotechnology)

HomeWork…


Read Pg 403
-
409


page 409 questions 1
-
2


page 413 questions 16
-
21


Entry Task 1/27…


How do police determine if a suspects
DNA is the same as the DNA found at a
crime scene?



Turn in Book Work…

Today…


Solving a Crime…


DNA Fingerprinting Article

Solving a Crime


With your table partner read “to catch a
thief”


Follow Directions to Solve the Crime

After you Solved the Crime…


Recycle DNA, leave instructions on table


Pick up article


Answer Questions on Same paper you
used for solving the crime questions



Don’t forget about the book work that was
assigned yesterday….did you turn it in?



Genetic Engineering


Genetic Engineering makes use of
recombination techniques which fuse
genes with host DNA (usually bacteria)


This allows large quantities of isolated
genes and their products (proteins) to be
made


Transformation of DNA places this new
foreign DNA into host organisms


Genetic Engineering


Recombinant DNA


A segment of DNA from organism 1 is
placed into the DNA from organism 2


This new (recombinant) DNA is then placed
into organism 2. This is called
Transformation


Organism 2 then produces the protein that
is usually produced by organism 1

Why is this beneficial?????

Genetic Engineering


Genetic engineering is making substantial
contributions to biological research, medicine,
industry, agriculture, and many other arenas


Human Insulin can now be produced by bacteria
in laboratories at a low cost


Gene therapy uses recombinant DNA to add a
missing gene to individuals. Results/Ethics???


Non
-
molecular genetic engineering
techniques have been in place for centuries
and are also of continued importance

PCR


Purpose: to make a huge number of
copies of a gene for sequencing or gel
analysis.


Consists of 3 major steps that are
repeated for 30


40 cycles

PCR


The Steps


Step 1


Denaturation


At 94
°
C the two strands of DNA separate


Step 2


Annealing


At ~ 55 C The Primers bind to the
complementary region of DNA strands


Step 3


Elongation


At ~ 72 C DNA Polymerase
(Taq Polymerase)

synthesizes a new DNA molecule


PCR


These three steps are repeated several
times


usually about 30 cycles


The target sequence of DNA is
amplified exponentially


30 cycles yield
about 1 billion copies


This Technique is used for many
different applications


research,
medicine, sequencing
-
criminal cases

1
st

step
-

Denaturation


94
°

C


Causes complementary DNA strands to
separate

2
nd

step
-

Annealing


54
°

C


Allows “primers” to join DNA strands


Primer


A sequence of about 20


30
bases that is complementary to the
beginning sequence of the target DNA
to be copied.

Primers


2 different primers are needed


one for each complementary side
of DNA.






5’ AG
CTTACT
TA 3’

3’ TC
GAATGA
AT 5’

Primer Question

If the red area is the target sequence,
what 2 primers would work for this
DNA?



5’ AG
CTTACT
TA 3’



3’ TC
GAATGA
AT 5’


a.
5’ CTT



c. 5’ AGT

b.
5’ GAA



d. 5’ TGA

3
rd

step
-

Extension


72
°

C


Uses an enzyme called polymerase


Polymerase takes the single bases and
builds complementary DNA from the
original DNA, beginning after the primer
on each strand

Polymerase


The polymerase is taken from a bacteria
that lives in hot springs,
Thermus
aquaticus
! Or
TAQ Polymerase


Unlike polymerase from other
organisms, this one will still work at hot
temperatures.

Repetition of 3 steps


30


40 cycles later, there are a billion
copies of the target DNA!


Animation





Get it or not


Stuff I think is Interesting


80
-
97% of Human DNA is non
-
coding


If you unraveled all of your DNA from
every cell, end to end, the strands would
stretch from the Earth to the Moon about
6,000 times


Genomically speaking, all races are equal,
you cannot tell simply by looking at
someone’s DNA what their race is

Stuff I think is Interesting

o
If you were to recite the order of ATCG’s in your
DNA at a rate of 100/min., 57 years would pass
before you reached the end (provided that you did
not stop to eat, drink, sleep, use the bathroom
etc.)

o
It would take a person typing 60 words per
minute, eight hours a day, around 50 years to
type the human genome.

o
You would need 3 gigabytes of storage space on a
computer to hold all of the information in your
genome


Stuff I think is Interesting

Organism


# of genes


Yeast


6,000


Drosophila


13,000


Nematode


18,000


Flowering plant

26,000


Human


30,000

Organism

# of chromosomes


Human


46


Gorilla


48


Dog



78


Dove


16


Butterflies


380


Bananas


66

o
Humans are 99.9%
genetically identical


only 0.1% of our
genetic make
-
up
differs

o
we share 98% of our
genes with
chimpanzees, 90%
with mice, 85% with
zebra fish, 21% with
worms