Biotechnology and Genomics

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1 Chapter14: pp BIOLOGY 10th Edition Biotechnology and Genomics Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. DNA probe array tagged DNA did bind to probe DNA probe Sylvia S. Mader tagged DNA tagged DNA did not bind to probe testing subject's DNA PowerPoint Lecture Slides are prepared by Dr. Isaac Barjis, Biology Instructor 1 Copyright The McGraw Hill Companies Inc. Permission required for reproduction or display

2 Outline DNA Cloning Recombinant DNA Technology Restriction Enzyme DNA Ligase Polymerase Chain Reaction Biotechnology Products Transgenic Bacteria, Plants, and Animals Gene Therapy Genomics 2

3 DNA Cloning Cloning is the production of identical copies of DNA Members of a bacterial colony on a petri dish are clones because they all came from division of the same cell. Identical twins are clones Single embryo separate to become two. Gene cloning is production of many identical copies of the same gene. If the inserted gene is replicated and expressed, we can recover the cloned gene or protein product. Cloned genes have many research purposes Humans can be treated with gene therapy 3

4 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at 4

5 Recombinant DNA Technology 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 Two enzymes are required to introduce foreign DNA into vector DNA A restriction enzyme - cleaves DNA, and A DNA ligase enzyme - seals DNA into an opening created by the restriction enzyme 5

6 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at

7 Cloning a Human Gene Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. human DNA plasmid human cell bacterium insulin gene 1. Restriction enzyme cleaves DNA. 2. DNA ligase seals human gene and plasmid. recombinant DNA 3. Host cell takes up recombined plasmid. 4. Gene cloning occurs. insulin 7

8 Restriction Enzyme Cuts DNA at specific points. Cleaves vector (plasmid) and foreign (human) DNA. Cleaving DNA makes DNA fragments ending in short single-stranded segments with sticky ends. The sticky ends allow insertion of foreign DNA into vector DNA. Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. DNA duplex A T G C A T A T T A T A C G G C C G restriction enzyme A A T T C G C A G "sticky ends" G C G T C T T A A 8

9 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at

10 DNA Ligase Seals the foreign gene into the vector DNA Treated cells (bacteria) take up plasmids Bacteria and plasmids reproduce. Many copies of the plasmid and many copies of the foreign gene. 10

11 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at

12 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at

13 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at

14 DNA Cloning: Polymerase Chain Reaction (PCR) Amplifies a targeted sequence of DNA Create millions of copies of a single gene or a specific piece of DNA in a test tube Requires: DNA polymerase Withstands the temperature necessary to separate double-stranded DNA. A supply of nucleotides for the new, complementary strand 14

15 PCR Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. PCR cycles DNA copies first 1 DNA double strand second 2 new strand old strand third 4 new old fourth 8 old new fifth 16 and so forth 15

16 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at 16

17 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at

18 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at

19 Applications of PCR: Analyzing DNA Segments DNA fingerprinting is the technique of using DNA fragment lengths 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. 19

20 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at

21 Fluorescence units Base repeat units DNA Fingerprinting & Paternity Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. few Mother Child Male 1 Male 2 many a. DNA Band patterns b. Automated DNA fingerprinting Increasing size 21

22 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at 22

23 Biotechnology Products Genetically engineered organisms can produce biotechnology products. Organisms that have had a foreign gene inserted into them are transgenic. 23

24 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at

25 Transgenic Bacteria Gene of interest is inserted into bacteria. Bacteria are grown in large vats called bioreactors and product is harvested. Products on the market include insulin, hepatitis B vaccine, t-pa, and human growth hormone. Transgenic bacteria can produce chemical products. Transgenic bacteria process minerals. Metals Collection 25

26 Transgenic Bacteria Transgenic bacteria promote plant health Bacteria that colonize corn roots can be endowed with genes for insect toxin Transgenic bacteria can degrade substances. Oil-Eating Bacteria 26

27 Genetically Engineered Bacteria Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. (Both): Courtesy General Electric Research & Development 27

28 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at 28

29 Transgenic Plants Agricultural Crops Foreign genes now give cotton, corn, and potato strains the ability to produce an insect toxin Soybeans are now resistant to a common herbicide Human Hormones Plants are being engineered to produce human proteins including hormones, clotting factors, and antibodies in their seeds 29

30 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at 30

31 Transgenic Animals Vortex Mixing: Many types of animal eggs have taken up the gene for bovine growth hormone (bgh) The procedure has been used to produce larger fishes, cows, pigs, rabbits, and sheep Gene Pharming: Use of transgenic farm animals to produce pharmaceuticals Genes coding for therapeutic & diagnostic proteins are incorporated into an animal s DNA The proteins appear in the animal s milk Plans are to produce drugs to treat Cystic fibrosis Cancer Blood diseases, etc. 31

32 Transgenic Mammals Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. human gene for growth hormone microinjection of human gene donor of egg development within a host goat human growth hormone Transgenic goat produces human growth hormone. milk a. transgenic goat cells with gene for human growth hormone microinjection of transgenic gene into enucleated donor eggs enucleated eggs donor of eggs development within host goats milk Cloned transgenic goats produce human growth hormone. b. 32

33 Transgenic Animals Researchers are using transgenic mice for various research projects. Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. one-celled mouse embryos with two X chromosomes no injection inject SRY DNA Embryo develops into a female. Embryo develops into a male. FEMALE MALE 33

34 Gene Therapy Gene therapy involves procedures to give patients healthy genes to make up for a faulty gene. It also includes the use of genes to treat genetic disorders and various human illnesses. There are ex vivo (outside body) and in vivo (inside body) methods of gene therapy. Ex Vivo Children with Severe Combined Immunodeficiency Bone Marrow Stem Cells In Vivo Cystic Fibrosis Nasal / Respiratory Spray 34

35 Gene Therapy Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Brain (gene transfer by injection)* Huntington disease Alzheimer disease Parkinson disease brain tumors Skin (gene transfer by modified blood cells)** skin cancer Lungs (gene transfer by aerosol spray)* cystic fibrosis hereditary emphysema Liver (gene transfer by modified implants)** familial hypercholesterolemia Blood (gene transfer by bone marrow transplant)* sickle-cell disease Endothelium (blood vessel lining) (gene transfer by implantation of modified implants)** hemophilia diabetes mellitus Muscle (gene transfer by injection)* Duchenne muscular dystrophy Bone marrow (gene transfer by implantation of modified stem cells)** SCID sickle-cell disease * in vivo ** ex vivo 35

36 Genomics Genomics is the study of genomes of humans and other organisms. Sequencing the Bases The Human Genome Project produced a working draft of all the base pairs in all chromosomes. Took 13 years to sequence three billion base pairs along the length of chromosomes. 36

37 Human Genome Project Genome - All the genetic information of an individual (or species) Goals of Human Genome Project Determine the base pair sequence Construct a map showing sequence of genes on specific chromosomes 37

38 Human Genome Project Humans have 20,500 genes Most code for proteins Much of the human genome was formerly described as junk Does not specify the order of amino acids in a polypeptide Recent observation suggest that between 74% and 93% of the genome is transcribed into RNA Thus, vast junk DNA wasteland may be much more important than once thought 38

39 Eukaryotic Gene Structure Historically, genes were defined as discrete units of heredity that corresponded to a locus on a chromosome. Prokaryotes typically possess a single circular chromosome. Eukaryotic chromosomes are much more complex. 39

40 Chromosomal DNA Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. exon exon intergenic sequences Gene C DNA intron Gene A Gene B pre-mrna exon intron exon RNA introns Gene A mrna Gene B mrna Gene C mrna Cindy Charles/PhotoEdit 40

41 Eukaryotic Gene Structure An Intergenic Sequences are DNA sequences that occur between genes Repetitive DNA elements occur when the same sequence of two or more nucleotides are repeated many times along the length of one or more chromosomes. Transposons are specific DNA sequences that have the remarkable ability to move within and between chromosomes. 41

42 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at

43 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at

44 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at

45 Genomic Differences Between Species Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. 45

46 HapMap Project People inherit patterns of sequence differences, called haplotypes If one haplotype of a person has an A rather than a G at a particular location in a chromosome, there are probably other particular base differences near the A Genetic data from African, Asian, and European populations will be analyzed A HapMap is a catalog common sequence differences that occur in a species The goal of the project is to link haplotypes to risk for specific illnesses May lead to new methods of preventing, diagnosing, and treating disease 46

47 Functional and Comparative Genomics Functional genomics aims to understand the role of genome in cells or organisms DNA microarrays can monitor the expression of thousands of genes simultaneously and tell us: What genes are turned on Environmental conditions that turn on the gene DNA microarrays contain microscopic amounts of known DNA fixed onto a small glass slide mrna bind through complementary base pairing mrna is produced by active cell. Identify various mutations in the genome of an individual This is called the person s genetic profile. 47

48 DNA Microarray Technology Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. DNA probe array tagged DNA did bind to probe DNA probe tagged DNA tagged DNA did not bind to probe testing subject's DNA 48

49 Animation Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the Normal or Slide Sorter views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at 49

50 Genetic Profile The complete genotype of an individual This is the person s genetic profile A way of studying how genes work together to control the phenotype Analyze the genetic profile of many individuals Compare their profiles to their phenotypes 50

51 Proteomics The study of the structure, function, and interaction of cellular proteins At least 25,000 of our genes are translated into proteins The sum total of these proteins is called the human proteome Understanding protein function is essential to the development of better drugs Correlate drug treatment to the particular genome Increase efficiency and decrease side effects Once the primary structure of these protein is known It should be possible to predict their tertiary structure Computer modeling of the tertiary of these proteins is an important part of proteomics 51

52 Bioinformatics The application of computer technologies to the study of the genome Genomics and proteomics produce raw data These fields depend on computer analysis to find significant patterns in the data Scientists hope to find relationships between genetic profiles and genetic disorders New computational tools will be needed to accomplish these goals 52

53 Bioinformatics Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. exon exon intergenic sequences Gene C DNA intron Gene A Gene B pre-mrna exon intron exon RNA introns Gene A mrna Gene B mrna Gene C mrna 53

54 Review DNA Cloning Recombinant DNA Technology Restriction Enzyme DNA Ligase Polymerase Chain Reaction Biotechnology Products Gene Therapy Genomics 54

55 Chapter14: pp BIOLOGY 10th Edition Biotechnology and Genomics Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. DNA probe array tagged DNA did bind to probe DNA probe Sylvia S. Mader tagged DNA tagged DNA did not bind to probe testing subject's DNA PowerPoint Lecture Slides are prepared by Dr. Isaac Barjis, Biology Instructor 55 Copyright The McGraw Hill Companies Inc. Permission required for reproduction or display

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