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UMass Amherst MICROBIO 310 - Transgenic and Genetically Engineered Plants and Animals

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Microbio 310 1st Edition Lecture 18 Outline of Last Lecture I. 13.10 Nitrification II. 13.11 AnammoxIII. 13.12 The Calvin CycleIV. 13.13 Other Autotrophic Pathways in PhototrophsV. 13.14 Nitrogenase and Nitrogen FixationVI. 13.15 Genetics and Regulation of Nitrogen FixationVII. 15.1 Industrial Products and the Microorganisms That Make ThemVIII. 15.2 Production and ScaleIX. 15.3 Antibiotics: Isolation, Yield, and PurificationX. 15.4 Industrial Production of Penicillins and TetracyclinesXI. 15.5 Vitamins and Amino AcidsOutline of Current Lecture I. 15.6 Enzymes as Industrial ProductsII. 15.9 BiofuelsIII. 15.10 Expressing Mammalian Genes in BacteriaIV. 15.11Production of Genetically Engineered SomatotropinV. 15.12 Human proteins from microbesVI. 15.16 Genetic Engineering of AnimalsThese notes represent a detailed interpretation of the professor’s lecture. GradeBuddy is best used as a supplement to your own notes, not as a substitute.VII. 15.17 Gene Therapy in HumansVIII. 15.18 Transgenic Plants in Agriculture Current Lecture15.6 Enzymes as Industrial Products• Exoenzymes– Enzymes that are excreted into the medium instead of being held within the cell; they are extracellular– Can digest/breakdown insoluble polymers such as cellulose, protein, and starch• Enzymes are useful as industrial catalysts – Produce only one stereoisomer– High substrate specificity• Enzymes are produced from fungi and bacteria– Bacterial proteases are used in laundry detergents (can also contain amylases, lipases, and reductases)• Isolated from alkaliphilic bacteria• Amylases and glucoamylases are also commercially important– Produce high-fructose syrup• Extremozymes– Enzymes that function at some environmental extreme (e.g., pH or temperature) – Ex: An acid-tolerant enzyme mixture is used as a feed supplement for poultry. The enzymes function in the bird’s stomach to digest fibrous materials in the feed, thereby improving the nutritional value of the feed and promoting more rapid growth. – Produced by extremophiles15.9 Biofuels• Ethanol Biofuels– Ethanol is a major industrial commodity chemical– Over 60 billion liters of alcohol are produced yearly from the fermentation of feedstocks Gasohol and E-85 • Petroleum Biofuels– Production of butanol– Synthesis of petroleum from green algae15.10 Expressing Mammalian Genes in Bacteria• Biotechnology– Use of living organisms for industrial or commercial applications• Genetically modified organism (GMO)– An organism whose genome has been altered• Genetic engineering allows expression of eukaryotic genes in prokaryotes (e.g., insulin)• This is achieved by:– Cloning the gene via mRNA– Finding the gene via the protein• Protein synthesis in a foreign host is subject to other problems– Degradation by intracellular (host cell) proteases – Eukaryotic proteins may be toxic to prokaryotic host, killing the host– Formation of inclusion bodies: aggregated insoluble proteins that are often misfolded or partly denatured and can be toxic to host (not bioactive protein)• Fusion of a target protein with a carrier protein facilitates protein purification15.11 Production of Genetically Engineered Somatotropin• Insulin was the first human protein made commercially by genetic engineering• Somatotropin, a growth hormone, is another widely produced hormone– Cloned as cDNA from the mRNA– Recombinant bovine somatotropin (rBST) is commonly used in the dairy industry; increases milk production in cows15.12 Human proteins from microbes• Erthropoetin RBC blood growth factor-treats anemia• TPA (tissue plasminogen activator)-used to prevent heart attacks by dissolving blood clots• DNAse Cystic Fibrosis biofilm- treats the build-up of DNA-containing mucus in CF patients• Interferon• Factor VIII-promotes blood clotting• rBGH• Insulin• Gardasil VLPs (virus-like particles)15.16 Genetic Engineering of Animals• Genetic engineering can be used to develop transgenic animals• Transgenic animals are useful for:– Producing human proteins that require specific post-translational modifications– Medical research– Improving livestock and other food animals for human consumption (ex: making salmon grow faster)15.17 Gene Therapy in Humans• Gene therapy holds promise for tackling many human genetic diseases• Gene therapy: introduces a functional copy of a gene to treat a disease caused by a dysfunctional version of the gene• The use of recombinant DNA technology and conventional genetic studies allows for the localization of particular genetic defects to specific regions of the genome15.18 Transgenic Plants in Agriculture• Plants can be genetically modified through several approaches, including:– Electroporation– Particle gun methods– Use of plasmids from bacterium Agrobacterium tumefaciens• Many successes in plant genetic engineering; several transgenic plants are in agricultural production• The plant pathogen Agrobacterium tumefaciens can be used to introduce DNA into plants• A. tumefaciens contains the Ti plasmid, which is responsible for virulence• The Ti plasmid contains genes that mobilize DNA for transfer to the plant– The segment of the Ti plasmid that is transferred to the plant is called the T-DNA• Tobacco was the first genetically modified (GM) plant to be grown commercially– 2005 estimate: >1 billion acres of agricultural land are used to grow GM crops• Several areas are targeted for genetic improvements in plants including herbicide, insect, and microbial disease resistance as well as improved product quality• Plants are engineered to have herbicide resistance to protect them from herbicides applied to kill weeds (e.g., glyphosate)• One of the most widely used approaches for genetically engineering insect resistance in plantsinvolves the introduction of genes encoding the toxic protein of Bacillus thuringiensis (Bt toxin)• Improving product quality is another target area of genetic engineering of plants– For example, spoilage delay• Transgenic plants can also be employed to produce human proteins for medical use– Examples: interferon, antibodies,


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UMass Amherst MICROBIO 310 - Transgenic and Genetically Engineered Plants and Animals

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