Bio Sci 97 Part 2 - Cloning Genes vs Cloning Organisms...

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Unformatted text preview: Cloning Genes vs Cloning Organisms Lectures Notes Recombinant Cloning Cloning Genes Insert a particular fragment of interest of DNA into a vector A vector is another DNA molecule that can be put into a host This creates a new DNA molecule Recombinant DNA A few of the many uses of Recombinant Technology Can produce many DNA copies of the cloned gene Can produce large quantities of a specific protein Food Tomatoes unsuccessful cloned the gene and made a reverse copy of it, put that into the tomato Knocked out the function of the enzyme that makes the tomato soft Did last longer, but fewer tomatoes Flavr-savr Beta-carotene golden rice successful Beta-carotene Makes carrots orange Precursor to vitamin A Took genes from rice Orange rice already have vitamin A present Manufacturing Medical Transgenic proteins like Factor VIII Hemophilia present in humans on X chromosome do not produce factor VIII no blood clotting Russian family death of hemophilia Produce factor VIII, and provide it to patients Using blood, but problems with purification of blood Purification not effective can cause hepatitis, kill patients Drug discovery Gene Therapy Biotech Structural biology (crystallography) Take a purified structure X-ray crystallography Molecule structure Transgenic Distinguish the modified organism Use PCR from tail Understanding gene function Rescue Biomarkers (Green Fluorescent Protein) Vectors Carrier of recombination Derived from plasmids Typically small (<10 Kb) (1 kB = 1000 base pairs) Not a chromosome but an extrachromosomal element such as a plasmid or a phage Plasmid must have single point of replication Phage prepare a vector that is more phage-like, and will make lots of copies of the snippet of DNA Can be transformed into a host cell (typically the bacterium E. coli ) Contains a replication origin, so it can replicate inside that host cell So we can make as many copies of the cloned gene as we need pBluescript plasmid origin of replication tells bacterial machinery the place to make copies from Selectable marker Produce cells in culture that dont do anything good Multiple cloning site polylinker place where DNA is actually inserted label for restriction endonuclease site each restriction site is unique in the plasmid Restriction Endonucleases and Restriction Sites : an enzyme (protein) that binds a specific (usually palindromic) DNA sequence function as a dimmer dimers come in and each cuts a strand 2 strands have to be complementary palindrome same forward as backward Restriction enzymes high degree of specificity Sticky ends area that was cut...
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This note was uploaded on 01/22/2012 for the course BIOLOGY SC Bio Sci 93 taught by Professor Dr.dianeo'dowdanddr.rahulwarrior during the Summer '10 term at UC Irvine.

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Bio Sci 97 Part 2 - Cloning Genes vs Cloning Organisms...

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