Genetic Technology Ch. 13

neighgreasycornerΒιοτεχνολογία

14 Δεκ 2012 (πριν από 4 χρόνια και 8 μήνες)

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Genetic Technology

DNA Identification


1
-
2% variation in human genome


Forensics, Military, Human Origins, Migration

http://www.youtube.com/watch?v=kEGfAubRogM

Steps in DNA identification

1)
Isolate the DNA in a sample (make copies)

2)
Cut DNA into shorter fragments

3)
Sort DNA via size

4)
Compare size fragments in the unknown
sample to those of known samples of DNA

http://www.youtube.com/watch?v=H_EwSCYuQ
bw


Polymerase Chain Reaction Machine


Polymerase Chain Reaction


Acquire a desired sample of DNA


Add DNA polymerase, free nucleotides,
primers, then heat it up!


DNA strands separate, cool the system to
allow primers to bind to original strands


DNA polymerase adds free nucleotides to
complete the strands (identical strands)


Heat again to repeat the process

DNA amplification via PCR

http://www.youtube.com/watch?v=eEcy9k_KsDI


Restriction Enzymes


An enzyme that destroys foreign DNA
molecules by cutting them at specific sites.

Cutting DNA via Restriction Enzymes


Restriction Enzymes: Sticky Ends



http://www.youtube.com/watch?v=
-
sI5vy
-
cD2g

Genetic Engineering/Recombinant
DNA


Genetic Engineering: A technology in which
the genome of a living cell is modified for
medical or industrial use



Recombinant DNA: DNA molecules that are
artificially created by combining DNA from
different sources

Genetic Engineering/Recombinant
DNA


Clones: Genes, Plants, Animals


Clone: an exact copy of a DNA segment, a
whole cell, or a complete organism.




Cloning the first humans


http://
www.youtube.com/watch?v=Tw1CX6ku0NQ




Cloning Via Vectors

1.
Isolate Plasmid and DNA of interest

2.
Cut both with a restriction enzyme

3.
Mix the DNA together and join them by using
DNA ligase

4.
Insert the recombinant plasmid into a host
bacterium

5.
Allow the bacterium to reproduce….What does
this do? Identify using a probe

http://www.youtube.com/watch?v=yta5KC18WkU
&feature=related


Human Genome Project


2% of the human genome codes for proteins


Chromosomes have unequal distribution of exons


30,000
-
40,000 protein coding genes


Exons are spliced in many different ways


½ the human genome arises from transposons
shuffling


8 million SNP’s
-

single nucleotide polymorphisms


J. Craig
Ventner


Creating Synthetic life part 1
-
6

http://www.youtube.com/watch?v=JdGRITKYZTQ&feat
ure=related

Gel Electrophoresis

1. Cut samples with a restriction enzyme

2. Run a current through the gel (negative goes to
positive electrode) smaller fragments go further

3. Transfer to nylon membrane, add a radioactive
probe

4. Expose X
-
Ray film/U.V. light to radiolabeled
membrane.



http://www.youtube.com/watch?v=QEG8dz7cbnY







1

2

3

4

5

6

7

8

Green

= Specific

primer’s
f
or plants
.


Red

=
Primer’s for a
specific
region common to GMO’s

Blue

= Blank Well

Black = Ladder


To Det
ermine Contents of
the Wells View Table
3A

500bp

200bp

Gene Therapy

1.
Isolate the functional gene


2. Insert the healthy gene into a viral vector


3.Introduce the recombinant virus to the patient
by infection (could be via nasal spray)

CFTR
-
cystic fibrosis transmembrane conductance
regulator
http
://
ghr.nlm.nih.gov/gene/CFTR

http
://learn.genetics.utah.edu/content/tech/genetherapy/cysticfibrosis/index.html


The CFTR gene provides instructions for making a protein called the cystic
fibrosis
trans membrane
conductance regulator. This protein functions as
a channel across the membrane of cells that produce mucus, sweat, saliva,
tears, and digestive enzymes. The channel transports negatively charged
particles called chloride ions into and out of cells. The transport of
chloride ions helps control the movement of water in tissues, which is
necessary for the production of thin, freely flowing mucus. Mucus is a
slippery substance that lubricates and protects the lining of the airways,
digestive system, reproductive system, and other organs and tissues.



The CFTR protein also regulates the function of other channels, such as
those that transport positively charged particles called sodium ions across
cell membranes. These channels are necessary for the normal function of
organs such as the lungs and pancreas.



CFTR continued


Disease
-
causing mutations in the CFTR gene alter the
production, structure, or stability of the chloride
channel. All of these changes prevent the channel
from functioning properly, which impairs the
transport of chloride ions and the movement of
water into and out of cells. As a result, cells that line
the passageways of the lungs, pancreas, and other
organs produce mucus that is abnormally thick and
sticky. The abnormal mucus obstructs the airways
and glands, leading to the characteristic signs and
symptoms of cystic fibrosis

MODEL ORGANISMS


Model Organisms


Genome is mapped


Used to understand human gene control


Induced mutations to find gene actions


Take info to make generalizations in larger
groups




Protein synthesis: an epic on the
cellular level


http://www.youtube.com/watch?v=u9dhO0iC
Lww