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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
• Nucleoid Region- is bacteria chromosome
• PLASMID- small rings of DNA that have only a few genes.
Plasmid
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/exhibitdna/index.jsp
Cloning
Cloning Activity
• http://gslc.genetics.utah.edu/units/cloning/clickandclone/
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