genetics notes 4

genetics notes 4 - DNA Technology 14:28 Golden Rice •...

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Unformatted text preview: DNA Technology 14:28 Golden Rice • Transgenic plant (genes from another organism incorporated into its cells) • Rice produces beta-carotene (vitamin A precursor- works with protein in eyes to help you see) o In areas where white rice is eaten a lot, blindness is a problem, scientists created this golden rice to help with that • Developed to help prevent blindness Genetically engineering rice • Infected them with agrobacterium, transferred 3 genes necessary to produce beta carotene into the rice plant’s cells, so now the when reproduce, have genes now o Two genes from a daffodil and one from a bacteria, each gene produces an enzyme along pathway for making beta carotene Recombinant DNA • The combination of fragments of DNA from different sources (cutting and pasting DNA fragments together) o How got the genes into the agrobacterium Cutting DNA • Restriction endonucleases (restriction enzymes) o Restricted to where it can cut based on the DNA sequence o Sticky ends – a region of single stranded DNA, some enzymes create this kind, preferable kind when wanting to paste DNA together, asymmetrical cut o Blunt ends – no single stranded region, symmetrical cut, harder to work with in the lab o Produced by bacteria • Nomenclature o Eco R1 recognizes AATT (palindromic sequence – most restriction enzymes are palindromic bc have to be able to cut both strands of DNA) o Can break phosphodiester bonds and hydrogen bonds How do we isolate/separate DNA fragments? • Say want the medium fragment – run gel electrophoresis • DNA migrates along an electric field by size and charge (negative bc of phospate groups) • Small fragments migrate towards bottom, bigger stays towards top • Can cut it out the gel • Then purify the DNA by heating Pasting DNA • Complementary single stranded ends bond • Then create new phosphodiester bonds using ligase Overview of Gene Cloning • Cutting and pasting is a part of this • Isolation of and copying of a particular gene • Isolate gene of interest, then put it in cloning vector (carries DNA of interest) (to make copies), cut it to make sticky ends, now have recombinant DNA molecule, put it in a bacteria, and the bacteria will replicate it for us whenever it divides Cloning vectors • Origin of replication – DNA polymerase can’t replicate it without it • Selectable marker o Antiobiotic resistance o Auxotrophic marker (nutrient marker) – doing selection based on a nutrient Ex yeast – whether could grow on –ade plate or not once plasmid inserted • Unique restriction sites o Where gene of interest is inserted o Aka multiple coning site (MSC) or polylinker...
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This note was uploaded on 06/23/2011 for the course BIO 212 taught by Professor Frederick during the Fall '07 term at Wofford.

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genetics notes 4 - DNA Technology 14:28 Golden Rice •...

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