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Biotechnology

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Biotechnology

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    1. Biotechnology Chapter 14

    4. 4 Recombinant DNA (rDNA) contains DNA from two or more different sources Requires: A vector introduces rDNA into host cell Plasmids (small accessory rings of DNA from bacteria) are common vectors Phage vectors (bacterial viruses) can also be used Two enzymes to introduce foreign DNA into vector DNA A restriction enzyme - cleaves DNA Bacterial enzyme that stops viral reproduction by cleaving viral DNA Act as molecular scisssors (cut plasmids and foreign human DNA) Produce short single stranded “sticky ends” where insertions of foreign DNA can be made A DNA ligase enzyme - seals DNA into an opening created by the restriction enzyme DNA Manipulation

    5. 5 Cloning a Human Gene

    6. 6 Genetic Engineering Has generated excitement and controversy Transgenic organisms have had a foreign gene inserted into their genetic make-up

    7. 7 Agricultural Applications Ti (tumor-inducing) plasmid is the most used vector for plant genetic engineering -Obtained from Agrobacterium tumefaciens, which normally infects broadleaf plants -However, bacterium does not infect corn, rice and wheat

    8. 8

    9. 9 Agricultural Applications Herbicide resistance -Broadleaf plants have been engineered to be resistant to the herbicide glyphosate -This allows for no-till planting

    10. 10 Agricultural Applications Pest resistance -Insecticidal proteins have been transferred into crop plants to make them pest-resistant -Bt toxin from Bacillus thuringiensis Golden rice -Rice that has been genetically modified to produce b-carotene (provitamin A) -Converted in the body to vitamin A

    11. 11 Agricultural Applications Adoption of genetically modified (GM) crops has been resisted in some areas because of questions about: -Crop safety for human consumption -Movement of genes into wild relatives -Loss of biodiversity

    12. 12 Agricultural Applications Biopharming -Transgenic plants are used to produce pharmaceuticals -Human serum albumin -Recombinant subunit vaccines -Against Norwalk and rabies viruses -Recombinant monoclonal antibodies -Against tooth decay-causing bacteria

    13. 13 Transgenic Mammals

    14. 14 The insertion of genetic material into human cells for the treatment of a disorder

    15. Recombinant DNA Vaccines?

    16. 16 Gene Therapy

    17. 17 DNA Analysis Polymerase chain reaction (PCR) -Developed by Kary Mullis (1983) (got Noblel) -Allows the amplification of a small DNA fragment using primers (supplied nucleotides) that flank the region -Allows thousands of copies to be made of small samples of DNA -Each PCR cycle involves three steps: 1. Denaturation (high temperature) 2. Annealing of primers (low temperature) 3. DNA synthesis (intermediate temperature) -Taq polymerase (DNA polymerase)

    18. 18 DNA Analysis Polymerase chain reaction (PCR) -Has revolutionized science and medicine because it allows the investigation of minute samples of DNA -Forensics -Detection of genetic defects in embryos -Analysis of mitochondrial DNA from early human species and other animals

    19. 19 PCR

    20. 20 Applications of PCR: Analyzing DNA Segments via Gel Electrophoresis DNA can be subjected to DNA fingerprinting Treat DNA segment with restriction enzymes A unique collection of different fragments is produced Gel electrophoresis separates the fragments according to their charge/size Produces distinctive banding pattern Usually used to measure number of repeats of short sequences Used in paternity suits, rape cases, corpse ID, etc.

    21. 21 Gel Electrophoresis A technique used to separate DNA fragments by size The gel (agarose or polyacrylamide) is subjected to an electrical field The DNA, which is negatively-charged, migrates towards the positive pole The larger the DNA fragment, the slower it will move through the gel matrix DNA is visualized using fluorescent dyes

    22. 22

    23. 23 DNA Fingerprinting & Paternity

    24. 24 DNA Analysis

    25. 25 Human Genome Project Genome - All the genetic information of an individual (or species) Goals of Human Genome Project (completed 2003) Determine the base pair sequence in all human DNA 13 year project Working draft Construct a map showing sequence of genes on specific chromosomes Other species in final stages

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