genetics_midterm3 - Genomic Library and cDNA Library differ...

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Genomic Library and cDNA Library differ in only the first two steps. The cDNA Library is very much smaller. The sequences are a lot different too. There are no introns, poly A tail or promotor. Applications of the two 1. Clone the gene so that you can express it to get the protein product in bacteria a. cDNA Library b. Bacteria can’t splice out introns, so cDNA is better because it doesn’t have introns. 2. Study the regulation of the gene a. Genomic Library b. Must have those regulatory sequences in tact 3. Splicing a. Genomic Library b. Must have introns Why do we clone? 1. To have an unlimited supply of the product that sequence encodes a. Industrial purposes b. Experimental purposes c. Vaccines d. Other pharmaceuticals 2. Advantages to doing it in bacteria a. Lower the cost and raise the efficiency of isolating the protein i. The isolation process is very easy and therefore much cheaper b. The bacteria get viruses through bacteriophages. The bacteria cannot carry human pathogens, so it’s far safer for the recipient. Gene Therapy 1. Genetic modifications in Humans 2. Done to Adults and older children 3. There are risks involved including cancer 4. People must understand the risks and be prepared for death 5. There is a target tissue 6. Use retroviruses and adenoviruses a. Replace pathogenic gene with normal gene for problem b. Problem is immune system fights virus infection 7. Using somatic cells 8. Not heritable Genetic Engineering 1. Genetic modifications in non-humans 2. Manipulating the plants and animals to help humans 3. Goal is to treat someone with a very serious disease and is often not curable 4. Done in embryos 5. Changes are not limited to somatic cells, but germ cells also 6. Changes are heritable and you want them to be so
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DNA Fingerprinting - Focus on the parts of the genome that vary greatly from person to person 1. Polymorphic areas 2. VNTR a. Variable Number of Tandem Repeats b. Need a quick and easy way to count the number of repeats at that location c. Want primers specific for the DNA just to either side of the repeats d. The sequences to which the primers attach are not polymorphic and therefore you can use the same primers for each person e. Do several rounds of PCR f. There are about a dozen or so regions that the FBI uses to make this “fingerprint” Mutation – Heritable Change in DNA There are many changes in DNA that aren’t heritable Lesion – Change in DNA that doesn’t create one of the 4 standard nucleotides. A lesion on the template strand will often cause DNA polymerase to put the wrong nucleotide on the other strand. So lesions lead to mutations. Mutagenesis - Where mutations come from - Lesion causes mutation Spontaneous mutations - Those that are not created by an outside agent acting on DNA - Arise out of chemical structure of DNA itself 1. Breakage of the bond between the base and the 1 prime carbon of Deoxyribose a. What still holds the sugar in place? b.
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genetics_midterm3 - Genomic Library and cDNA Library differ...

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