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Recombinant DNA Technology

Recombinant DNA Technology. aka gene splicing and genetic engineering. http://www.2dayblog.com/images/2007/september/glofish.jpg. Zebrafish (boring):. Jellyfish with GFP (protein):. Recombinant DNA technology isolates the GFP gene and clones it. A bacterium (prokaryote):.

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Recombinant DNA Technology

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  1. Recombinant DNA Technology aka gene splicing and genetic engineering http://www.2dayblog.com/images/2007/september/glofish.jpg

  2. Zebrafish (boring): Jellyfish with GFP (protein): Recombinant DNA technology isolates the GFP gene and clones it A bacterium (prokaryote): The bacteria reproduce in a petri dish e. coli http://home.biotec.or.th/NewsCenter/my_documents/my_pictures/F7AFD_plasmid.jpg http://www.postech.edu/~hjcha/jelyfish.jpg http://michaelscomments.files.wordpress.com/2007/12/zebra-fish-61290.jpg

  3. Plasmids Plasmids are circular DNA molecules containing genes usually for resistance to bacteriophages (viruses): Restriction Enzymes (Restriction endonucleases) are proteins that cut DNA at specific places…. ….called restriction sites: 4-8 nucleotide sequence that the enzyme “recognizes”

  4. Restriction Enzymes In jellyfish DNA: DNA is cut into many fragments (one of them will contain the GFP gene) Staggered: “sticky end” In plasmid: you want only one restriction site so it opens in only one place DNA ligase enzyme catalyzes covalent bonds in sugar-phosphate backbone (makes it permanent)

  5. Cloning The recombinant plasmid is reintroduced to the bacterium with new genes The process with which this happens is transformation The plasmid is called the vector Bacteria without new plasmids expresses gene X Nonrecombinant plasmids (rejoined) Bacteria with new plasmids No GFP gene Recombinant plasmids virus-resistant gene GFP gene restriction site within gene X (cuts it) glow!

  6. Uses of Recomb. DN. Tech. • Modification of living organisms • crops • gene therapy, human engineering • Not wanting to pass on diseases • GloFish • Large-scale production (cloning) • Insulin (1978 Nobel Prize in Medicine for restriction enzymes) • Sequencing • Human Genome Project • evolution can be traced between species

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