Biotechnology

sidewalkhallΒιοτεχνολογία

23 Οκτ 2013 (πριν από 4 χρόνια και 18 μέρες)

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Biotechnology

Chapter 20

Biotechnology


Genetic
Engineering


Recombinant
DNA

DNA Cloning


Plasmids

DNA Cloning


Gene Cloning


Allows
amplification
of a gene or
harvesting of
its subsequent
protein


DNA Cloning


Restriction Enzymes


Protect cell by cutting
up foreign DNA


Restriction Site


Bacterial DNA
protected by
methylation of A and
C bases


Restriction Fragments


Sticky End


Can be made
permanent with
DNA ligase



DNA Cloning


Cloning Vector


Genomic
Library


cDNA Library


DNA Cloning


Nucleic Acid
Hybridization


Nucleic Acid
Probe


Polymerase Chain Reaction (PCR)


Denaturation


Annealing


Extension


Each cycle
results in 2
n

yield of DNA
molecules

Gel Electrophoresis


Restriction
Fragment Length
Polymorphism
(RFLP “Rif
-
Lip”)


Southern Blotting

Gel Electrophoresis


Northern Blotting


Reverse Transcriptase Polymerase
Chain Reaction (RT
-
PCR)


DNA Microarray

Determining Gene Function


In vitro mutagenesis


RNA interference
(RNAi)


Genome Wide
Association Studies


Single Nucleotide
Polymorphism (SNP)
“snip”


If SNP and disease
causing allele are
close, not often
separated by
crossing over

Organismal Cloning


Totipotent


Nuclear Transplantation


Ex. Dolly the Sheep, CC
the cat


Problems arise due to
epigenetic changes
causing improper gene
expression during
development

Organismal Cloning


Stem Cells


Embryonic Stem
(ES) Cells


Pluripotent


Therapeutic
cloning


Induced
pluripotent stem
(iPS) cells


Gene Therapy


Genetic Profile


Short Tandem Repeats (STRs)


# of repeats at different loci can be
used to compare individuals and
samples (forensics)

Safety and Ethical Questions about
Genetically Modified Organisms