The-Science-of-Transgenics

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Slide 1: 

The Science of Transgenics Phil McClean Department of Plant Science North Dakota State University The Sociological, Economic, and Ethical Impact of Transgenic Organisms Workshop Fargo, ND February 21, 2003

Slide 2: 

Transgenics are a Biotechnology Product How about some definitions Biotechnology - General Definition The application of technology to improve a biological organism Biotechnology - Detailed Definition The application of the technology to improve the biological function of an organism by adding genes from another organisms

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What About the Term Genetic Engineering? Genetic engineering involves: Isolating genes Modifying genes so they function better Preparing genes to be inserted into a new species Developing transgenes Genetic engineering is the basic tool set of biotechnology

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What is a transgenic? Transgene – the genetically engineered gene added to a species Ex. – modified EPSP synthase gene (encodes a protein that functions even when plant is treated with Roundup) Transgenic – an organism containing a transgene introduced by technological (not breeding) methods Ex. – Roundup Ready Crops Concept Based on the Term Transgene

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We can develop organisms that express a “novel” trait not normally found in the species Why are transgenics important? Extended shelf-life tomato (Flavr-Savr) Herbicide resistant soybean (Roundup Ready)

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Agriculture Transgenics On the Market Source: USDA Insect resistant cotton – Bt toxin kills the cotton boll worm transgene = Bt protein Insect resistant corn – Bt toxin kills the European corn borer transgene = Bt protein Normal Transgenic

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Virus resistance - papya resistant to papaya ringspot virus transgene = virus coat protein Source: Monsanto Herbicide resistant crops Now: soybean, corn, canola Coming: sugarbeet, lettuce, strawberry alfalfa, potato, wheat (2005?) transgene = modified EPSP synthase or phosphinothricin-N-acetyltransferase

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Biotech chymosin; the enzyme used to curdle milk products transgene = genetically engineered enzyme bST; bovin somatotropin; used to increase milk production transgene = genetically engineered enzyme Source: Rent Mother Nature Source: Chr. Hansen

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Some Ag Biotech Products Are Discontinued Poor Quality FlavrSavr tomatoes (Calgene) Negative Consumer Response Tomato paste (Zeneca) Negative Corporate Response NewLeaf (Monsanto) Universal Negative Publicity StarLink corn (Aventis) Why???

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Next Generation of Ag Biotech Products Source: Minnesota Microscopy Society Golden Rice – increased Vitamin A content (but not without controversy) transgene = three pathway enzymes Sunflower – white mold resistance transgene = oxalate oxidase from wheat

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Turfgrass – herbicide resistance; slower growing (= reduced mowing) Bio Steel – spider silk expressed in goats; used to make soft-body bullet proof vests (Nexia)

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Biotechnology is Not Just on the Farm Disease Treatment Diagnostics   Environmental Cleanup   Human Applications

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Human Applications Pharmaceutical products New solutions to old problems   Disease diagnosis Determine what disease you have or may get  Gene therapy Correcting disease by introducing a corrective gene

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Biotechnology and Health The genes for these proteins are: Cloned Inserted into bacteria Product isolated using biofermentation

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Environmental Applications Bioremediation - cleanup contaminated sites; uses microbes designed to degrade the pollution Indicator bacteria – contamination can be detected in the environment

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Tooth decay – engineered Streptococcus mutans, the bacteria that destroys enamel Future Health-related Biotech Products Vaccines – herpes, hepatitis C, AIDS, malaria

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Edible Vaccines Transgenic Plants Serving Human Health Needs Works like any vaccine A transgenic plant with a pathogen protein gene is developed Potato, banana, and tomato are targets Humans eat the plant The body produces antibodies against pathogen protein Humans are “immunized” against the pathogen Examples: Diarrhea Hepatitis B Measles

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A Popular Term We Need To Know GMOs - Genetically modified organisms GMO - an organism that expresses traits that result from the introduction of foreign DNA Originally a term equivalent to transgenic organism

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Some claim any improved biological product is a GMO They feel this will For example, some call plant varieties biotechnology products This is a false claim The GMO Ruse ease the publics fear pave the way for product acceptance

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Let’s Be Up Front Biotechnology adds traits not available in the species  Soybean does not have a gene to breakdown Roundup The gene comes from bacteria Breeding ? Biotechnology  Breeding only exchanges genes found in the species Breeding can transfer the transgene to other breeding materials  BUT this does not make it a biotechnology procedure

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Important Plant Improvement Methods Breeding Crossing two individuals from the same species; produces a new, improved variety; not a biotechnology procedure Transformation Adding a gene from another species; the essential biotechnology procedure to produce transgenics Source: USDA Source: USDA

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Wheat Rye Triticale X Interspecific Cross New species, but NOT biotechnology products

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Mutagenesis A useful procedure to produce a new trait But the normal gene is modified A transgene is not involved The product of mutagenesis is not a GMO

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ATTCGA ATTGGA Susceptible Normal Gene Resistant Mutant Gene Mutagenesis Treatment Mutagenesis Changes the DNA Sequence

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BASF Clearfield Products Herbicide resistance imidazolinones Mutant AHAS enzyme developed by mutagenesis Crops Canola Corn Rice Sunflower Wheat A Major Marketing Advantage but lost when stacked with a transgene

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The Roundup Ready Story Glyphosate is a broad-spectrum herbicide Active ingredient in Roundup herbicide Kills all plants it come in contact with Inhibits a key enzyme (EPSP synthase) in an amino acid pathway Plants die because they lack the key amino acids A resistant EPSP synthase gene allows crops to survive spraying

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+ Glyphosate X Roundup Sensitive Plants X X Without amino acids, plant dies X

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Bacterial EPSP synthase Shikimic acid + Phosphoenol pyruvate 3-enolpyruvyl shikimic acid-5-phosphate (EPSP) Aromatic amino acids Roundup Resistant Plants + Glyphosate With amino acids, plant lives RoundUp has no effect; enzyme is resistant to herbicide

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The Golden Rice Story Vitamin A deficiency is a major health problem Causes blindness Influences severity of diarrhea, measles >100 million children suffer from the problem For many countries, the infrastructure doesn’t exist to deliver vitamin pills Improved vitamin A content in widely consumed crops an attractive alternative

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?-Carotene Pathway Problem in Plants

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The Golden Rice Solution Daffodil gene Single bacterial gene; performs both functions Daffodil gene ?-Carotene Pathway Genes Added

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Introducing the Gene or Developing Transgenics Steps 1. Create transformation cassette 2. Introduce and select for transformants

Slide 33: 

Transformation Cassettes Contains

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Transformation Steps Prepare tissue for transformation Introduce DNA Culture plant tissue Field test the plants Agrobacterium or gene gun Multiple sites, multiple years

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Transformation cassettes are developed in the lab They are then introduced into a plant Two major delivery methods Delivering the Gene to the Plant

Slide 36: 

The Lab Steps

Slide 37: 

The Next Test Is The Field Herbicide Resistance

Slide 38: 

Final Test of the Transgenic Consumer Acceptance RoundUp Ready Corn