Genetically Modified Foods

The essential questions?

What ethical issues arise from genomic manipulation?

What are the societal implications?

How do scientist manipulate DNA and the genome of an organism?

Project ARISE: Advancing Rhode Island Science Education Funding provided by a Science Education Partnership Award from the National Center for Research Resources

Genetically Modified Foods

Student will know:
Students who complete this unit should have a better understanding of the technology used to develop GM foods and any potential risks and benefits of genetically modifying organisms.

Project ARISE: Advancing Rhode Island Science Education Funding provided by a Science Education Partnership Award from the National Center for Research Resources

Genetically Modified Foods

Students should ask:
Do we have enough information on GM foods to make an informed decision to support or reject GM foods?

Project ARISE: Advancing Rhode Island Science Education Funding provided by a Science Education Partnership Award from the National Center for Research Resources

Genetically Modified Food

ARISE August 3, 2009

Outline
How to make a GM organism Techniques Homework for tonight

Have you ever eaten genetically modified food?

Can you tell the difference between a genetically modified organism and a nonGM organism? Do GM foods taste any different? Could they?

http://www2.cast.ilstu.edu/ksmick/250/250lablink.htm

What is genetic modification?

Does genetic modification only happen in plants?
No, the first gene was transferred into bacteria.

What are some reasons for genetic modification?

Express recombinant insulin in bacteria

What are some of the benefits and some of the disadvantages of GM foods?
http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/A/Avery.html http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/R/RecombinantDNA.html#cloning

How long have humans been genetically modifying organisms?

What about in the lab? How long have scientists been modifying organisms? How is modern technology used to genetically modify organisms?

Teosinte

Why would we want to modify an organism?

Better crop yield, especially under harsh conditions Herbicide or disease resistance Nutrition or pharmaceuticals, vaccine delivery In 2004, approximately 85% of soy and 45% of corn grown in the U.S. were grown from Roundup Ready seed.
http://www.oercommons.org/courses/detecting-genetically-modified-food-by-pcr/

Roundup Ready Gene

The glyphosate resistance gene protects food plants against the broad-spectrum herbicide Roundup, which efficiently kills invasive weeds in the field. The major advantages of the "Roundup Ready system include better weed control, reduction of crop injury, higher yield, and lower environmental impact than traditional weed control systems. Notably, fields treated with Roundup require less tilling; this preserves soil fertility by lessening soil run-off and oxidation.

How to make a GM organism

Clone gene into vector (i.e. plasmid) with restriction enzymes and other molecular techniques Transform into organism or into biological vector (agrobacteria or virus) Infect plant with bacteria Select for transformants with herbicide

http://www.pbs.org/wgbh/harvest/

What we are doing today

Extract DNA from plant or food product Use the technique of PCR to copy a region of DNA found in Round-Up Ready foods Tomorrow we will analyze these products with gel elecrophoresis

Many of the same techniques are used to make a genetic modifications as to detect one
Polymerase Chain Reaction (PCR) Restriction enzymes Gel electrophoresis Transformation

PCR
Invented in 1983 by Kary Mullis (Nobel Prize in 1993 for its discovery) Uses primers to exponentially amplify a specific region of DNA Components needed for the reaction:
DNA Primers to region of interest DNA polymerase (Taq used to synthesize the DNA) dNTPS (the building blocks of the copied DNA) Buffer (with appropriate salts to ensure the enzyme works properly)

PCR
Three steps of the reaction:
Denaturation: High heat (94-98o) to separate the strands of DNA Annealing: (50-60o depends on the primers) this step allows the primers to bind to the denatured DNA strands Elongation (74o) DNA polymerase synthesizes the new strand
This step is dependant on the length of the product to be amplified (1min/1kb of DNA)

Check products with gel electrophoresis and sequencing

PCR: Cycles

PCR:

PCR: Thermocycler

PCR: Applications
Used to test for gene products for disease diagnosis Used to amplify small amounts of material
Forensics Fossil Records

Used for recombinant DNA technology Used for virus detection

Resources
Potential Problems
No amplification due to wrong buffer conditions No amplification due to lost enzyme activity Primers are wrong

Online Resources: http://www.dnai.org/b/index.html http://www.genome.gov/10000202 Click on Techniques

PCR is found in amplifying

Restriction Enzymes

Restriction Enzymes
Restriction enzymes are also called restriction endonucleases
They cut double stranded DNA at sequence specific sites They can produce sticky ends or blunt ends depending on the enzyme

Sticky Ends Blunt Ends Sticky Ends Blunt Ends

Restriction Enzymes
1978 Nobel Prize in Medicine was awarded to Daniel Nathans and Hamilton Smith for the discovery of restriction endonucleases
Restriction enzymes were discovered in E.coli as a defense mechanism against bacterial viruses (bacteriophages)

The recognition sites are usually 4-12 nucleotides long

Sequences are palindromic (GAATTC)

There are hundreds of restriction enzymes currently being used

Restriction Enzymes
What is better for making recombinant DNA: Sticky ends or blunt ends?

Restriction Enzymes
Student activity Potential Problems:
Wrong buffer * activity

Online resources http://www.dnai.org/b/index.html Click on Techniques

Restriction enzymes are found in cutting and pasting

Gel Electrophoresis
Gel electrophoresis is used to separate nucleic acids (DNA and RNA) or proteins for analytical use
DNA and RNA are separated using agarose Proteins are separated using polyacrylamide The gel is a matrix (cross-linked polymers) that allow products to be separated

Separation is based on the size (based on charge) of a product as it moves through a charged field

Gel Electrophoresis
The negative charge is at the top (closest to the samples) and the positive charge is at the bottom
Samples are negatively charged and will travel towards the positive charge
DNA and RNA are negative because of their sugarphosphate backbone Proteins are denatured to give a constant shape and given a charge through the negative loading buffer used

Samples are diluted in a loading buffer that helps the samples stay in the wells

Gel Electrophoresis
Applications Separating restriction digests Analyzing/purifying PCR products Sequencing Protein analysis

Gel Electrophoresis

Gel Electrophoresis

Gel Electrophoresis

Sample agarose gel stained with ethidium bromide (EtBr)

Gel Electrophoresis
Student activity Practice loading a gel with 20uL Kool Aid or food coloring Run gel and see color separation Discuss what it means for the colors to separate

Gel Electrophoresis

Gel Electrophoresis
Potential Problems Connecting the charges backward Not enough loading dye Running the gel too hot Handling EtBr

Online Resources: http://www.dnai.org/b/index.html Click on Techniques

Gel electrophoresis is found in sorting and sequencing

Homework
Activate your Brown ID: http://activate.brown.edu Read Teaching High School Science Chapters 1 and 2 Read GM foods & Teaching Critical Thinking Read GM Foods Unit and list three concerns or questions regarding the unit.

Restriction Enzymes

Restriction Enzymes: Applications

Restriction enzymes are commonly used in laboratories to create recombinant DNA Harvest DNA products for other applications DNase a general nuclease used to eliminate DNA in RNA samples

Gel Electrophoresis