Biotechnology
Biology- Regular
John Murnan
Etowah High School
Welcome to Biotech Inc.
Your new job requires some tools
• Biotech Inc. does research into developing
products for Genetic Engineering, DNA
fingerprinting and Cloning
• As interns you will learn how to use some
new tools and techniques
• Please open your tool box and lets see
what’s inside it.
Restriction Enzymes
– Act like scissors
– Cut DNA at specific places
Recognition sites of different
restriction enzymes
Restriction site function
Function
of
restriction
enzymes
Ligase
• Acts like glue
• Connects two pieces of DNA
– Makes bond between sugar-phosphate on
backbone.
DNA Polymerase
• Acts like a copier
– Copies a strand of DNA
• Requires free nucleotides to make the copies
Retroviruses
• Can insert DNA into a
chromosome in a
living cell
– HIV, the virus that
causes AIDS is one
type.
Some smaller tools
• Very thin syringe
– Used to remove nucleus from cells
• Micropipette
– A very small eyedropper
– Used to move very small amounts of fluid
Gel Electrophoresis
• Sorts molecules by size
– Uses a gel like jello and an electric current.
Gel Electrophoresis Setup
Gel
Electrophoresis
Pictures of gel electrophoresis
PCR
• Uses DNA Polymerase and free nucleotides
to make millions of copies of DNA from a
single strand.
Overview
of PCR
Effect of
PCR
Genetic Engineering
Manipulating organisms to create
products
• Use restriction enzymes, ligase, Gel
Electrophoresis and PCR
• Inserts foreign genes into bacteria
– Makes bacteria produce proteins
– Bacteria can be used to store genes
Plasmid Ligation
• Puts DNA from humans (or other species)
into bacteria DNA.
Review of Bacteria Structure
Plasmid
• Nucleoid Regionis bacteria
chromosome
• PLASMID- small
rings of DNA that
have only a few
genes.
Ligation of plasmid 1
Ligation of
plasmid 2
Genetic
Engineering
with plasmid
Possible
products of
genetic
engineering
Using Plasmid Ligation
techniques, we can make both
proteins and genes that can be
used to
DNA Fingerprinting
Forensic Scientists
• Use restriction enzymes, Gel
Electrophoresis, PCR
• Can identify any person based on their DNA
sequences.
• Each of us has unique sequences.
What is an RFLP?
Sometimes
mutations occur
which cause
recognition sites to
be changed.
If a person has a
mutation, then the
site is changed and
DNA of different
lengths is produced
using the same
restriction enzyme.
Restriction Fragment Length Polymorphism
Why you are different than other
people
Southern
Blot 1
Southern blot results
DNA
fingerprinting
example
• http://www.koshlandsciencemuseum.org/ex
hibitdna/index.jsp
Cloning
Cloning Activity
• http://gslc.genetics.utah.edu/units/cloning/cl
ickandclone/
Ethics of cloning
• Occurs naturallly
(identical twins)
• Easier screening of
embryos for genetic
diseases
• Increase chances for
infertile couples to
have children
• High risk of
developmental errors
leading to
miscarriages
• Cells are as old as the
individual they are
taken from
Gene Therapy
Gene
therapy
example
Gene
therapy
vectors
Delivery
methods
Gene Therapy
example