Session 16 (Recombinant DNA and Biotechnology)

Session 16 (Recombinant DNA and Biotechnology) - MCB 181...

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MCB 181 Study Session 16 (Recombinant DNA and Biotechnology)
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Learning Goals for Study Session 16 (Recombinant DNA and Biotechnology) Be able to clearly define the terms recombinant DNA, biotechnology, and genetic engineering. Describe the importance of restriction enzymes and the sticky ends they create in a DNA molecule. Briefly describe how a bacterial plasmid can be used to clone DNA and to select just those bacterial cells that have taken up a plasmid that contains the gene of interest. Be able to distinguish between genomic and cDNA libraries and the fundamental differences in the nature of the DNA clones in the two libraries. Briefly describe and state the uses for the PCR, dideoxy chain termination, gel electrophoresis, and Southern blot procedures. Be able to list and briefly describe some of the procedures for inserting genes into cells of organisms. Be able to list some of the ways in which DNA technology has been used to improve the lives of humans.
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Recombinant DNA and Biotechnology Recombinant DNA is DNA in which nucleotide sequences from two different sources (even different species) are combined in the laboratory to produce a new combination of genes. Biotechnology refers to the manipulation of organisms or their components to make novel and useful products. In this study session the laboratory methods that are used for making recombinant DNA are described. We also consider how some genetic engineering techniques have been applied to achieve breakthroughs in the field of biotechnology. We begin by considering several laboratory techniques that are essential for the manipulation of DNA.
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Restriction enzymes cut DNA molecules at specific nucleotide sequences! Restriction endonucleases are enzymes used by bacteria to defend themselves against intruding DNA from viruses or other organisms. Hundreds of different restriction enzymes have been discovered and characterized. These enzymes recognize a specific nucleotide sequence called the restriction site and cut both DNA strands as shown to the right. Notice that restriction sites are typically symmetrical with the same sequence on both strands in the 5’to 3’ direction and the cut is made in a staggered manner leaving sticky ends on each side of the cut. These sticky ends can hydrogen bond to any other DNA molecule cut with the same enzyme. Scientists use a DNA ligase enzyme to permanently join the DNA from the two sources producing a recombinant DNA molecule.
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DNA can be cloned into a bacterial plasmid to make many copies of the DNA molecule! Bacterial plasmids can be engineered to carry two genes, one for antibiotic (ampicillin) resistance and the second for production of β-galactosidase enzyme which can hydrolyze a special reagent to yield a blue color. The
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This note was uploaded on 03/24/2010 for the course MCB 181 taught by Professor Jorstad during the Spring '07 term at University of Arizona- Tucson.

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Session 16 (Recombinant DNA and Biotechnology) - MCB 181...

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