17btec - Remove the Gene Next, we must cut the gene out of...

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Lecture 17: Biotechnology Chapter 21: Applications
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The Big Picture What is biotechnology ? Real-life applications of scientific concepts
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Biotechnology Technology based on biology , agriculture, food science, or medicine Including genetic engineering
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Transgenic Organisms Transgenic bacteria, fungi, plants, animals Genetically modified by scientists for a specific purpose
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Genetic Modification Removing a gene from the DNA of one organism and inserting it into a different organism Recall: a gene is a sequence of DNA nucleotides that codes for a protein
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How Do We Do It? Gene cloning causes an organism to produce many copies of the same gene/protein But inserting the desired gene into the host organism requires a vector
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Vectors Molecule of DNA used to transfer genetic information Usually a plasmid Circular ring of accessory DNA found in bacteria
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Steps in Cloning a Gene 1.) Isolate the gene 2.) Remove the gene 3.) Re-insert the gene
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1.) Isolate the Gene Determine the order and location of nucleotides of the desired gene Radiolabel each of the bases and examine the order in which they appear
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2.)
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Unformatted text preview: Remove the Gene Next, we must cut the gene out of the rest of the DNA Using restriction enzymes Restriction Enzymes Cut DNA at a specific sequence E.g. EcoR1 cuts at AATT Use of Vectors Bacterial cells are able to take up plasmids from the surrounding environment We need to put our gene on a plasmid vector for transport Restriction Enzymes Again If we cut a plasmid with the same restriction enzyme that we used to remove the gene from the original organism the ends will match and can stick together Sealed by DNA ligase 3.) Re-insert the Gene Lastly, we have to insert the gene into our chosen cloning organism Usually bacteria (easy to manipulate) Chemical- and Heat-Treating Bacteria must be warmed in the presence of calcium chloride in order to take up the recombinant plasmids Exactly how they do this is a mystery Finally Cloning When the bacteria have taken up the plasmid , it can finally be cloned One bacterium makes multiple copies As they divide, the new bacterial cells also make multiple copies...
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This note was uploaded on 12/08/2011 for the course PRE-HEALTH 1160 taught by Professor L.miani during the Spring '11 term at Niagara College.

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17btec - Remove the Gene Next, we must cut the gene out of...

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