Lec3W11 - DNA Cloning If you introduce a foreign piece of...

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DNA Cloning If you introduce a foreign piece of DNA into a another organism, and that piece of DNA gets copied when the host cells replicate, you will have a large number of cells all with identical copies of that piece of foreign DNA – so you have “cloned” that piece of DNA, and can use the host cells to replicate it. If that piece of DNA codes for a protein, it is also possible for the host cells to produce the protein.
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Why clone DNA? Produce human proteins to use as drugs – insulin, growth hormone Make genetically modified plants that are resistant to pests or produce vitamins, etc Gene therapy – transfer good copy of a gene into a patient with abnormal copy Sequence genomes
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Figure 1.1 The basic steps in gene cloning. Gene Cloning and DNA Analysis by T.A. Brown. © 2006 T.A. Brown.
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What do you need to clone DNA? Source of DNA to be cloned genomic DNA or cDNA (DNA copied from RNA) Vector to get foreign DNA into host organism Plasmid, virus, phage, etc Type of vector you use depends on host Host organism Can be bacteria, yeast, mammalian cell, etc Bacteria easiest to use but there are issues with expressing eukaryotic proteins in bacteria
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Creating a genomic library A Genomic library is a collection of clones representing the entire genome of an organism 1. Isolate genomic DNA from source 2. Break up large pieces of chromosomal DNA into smaller pieces – vectors have limits in terms of sizes of insert
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This note was uploaded on 04/10/2011 for the course BIMM 101 taught by Professor Butler during the Spring '08 term at UCSD.

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Lec3W11 - DNA Cloning If you introduce a foreign piece of...

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