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Download CH 13 Gene Technoogy PPT
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GENE TECHNOLOGY Chapter 13 Human Genome Sequencing of the human genome was completed by 2007 Create a map showing where genes are located on human chromosomes DNA sequencing has depended on advances in technology, starting with making recombinant DNA In recombinant DNA, nucleotide sequences from two different sources, often two species, are combined in vitro into the same DNA molecule Using Restriction Enzymes to Make Recombinant DNA Bacterial restriction enzymes cut DNA molecules at specific DNA sequences called restriction sites A restriction enzyme usually makes many cuts, yielding restriction fragments The most useful restriction enzymes cut DNA in a staggered way, producing fragments with “sticky ends” that bond with complementary sticky ends of other fragments DNA ligase is an enzyme that seals the bonds between restriction fragments Fig. 20-2a Cell containing gene of interest Bacterium 1 Gene inserted into plasmid Bacterial chromosome Plasmid Recombinant DNA (plasmid) Gene of interest 2 2 Plasmid put into bacterial cell Recombinant bacterium DNA of chromosome Fig. 20-2b Recombinant bacterium 3 Host cell grown in culture to form a clone of cells containing the “cloned” gene of interest Protein expressed by gene of interest Gene of Interest Copies of gene Protein harvested 4 Basic research and Basic research on gene Gene for pest resistance inserted into plants various applications Gene used to alter bacteria for cleaning up toxic waste Protein dissolves blood clots in heart attack therapy Basic research on protein Human growth hormone treats stunted growth Fig. 20-2 Cell containing gene of interest Bacterium 1 Gene inserted into plasmid Bacterial chromosome Recombinant DNA (plasmid) Being able to genetically engineer bacteria for crops help keep cost low. Growing nonvirulent bacteria to clean oil spills is cost effective and a safer choice for our environment. Plasmid Gene of interest DNA of chromosome 2 Plasmid put into bacterial cell Recombinant bacterium 3 Host cell grown in culture to form a clone of cells containing the “cloned” gene of interest Gene of Interest Protein expressed by gene of interest Copies of gene Basic Protein harvested 4 Basic research and various applications research on gene Gene for pest resistance inserted into plants Gene used to alter bacteria for cleaning up toxic waste Protein dissolves blood clots in heart attack therapy Basic research on protein Human growth hormone treats stunted growth Genetically engineered proteins could mean less invasive procedures. An alternative to surgery. Fig. 20-3-1 Restriction site DNA 1 5 3 3 5 Restriction enzyme cuts sugar-phosphate backbones. Sticky end Fig. 20-3-2 Restriction site DNA 1 5 3 3 5 Restriction enzyme cuts sugar-phosphate backbones. Sticky end 2 DNA fragment added from another molecule cut by same enzyme. Base pairing occurs. One possible combination Fig. 20-3-3 Restriction site DNA 1 5 3 3 5 Restriction enzyme cuts sugar-phosphate backbones. Sticky end 2 DNA fragment added from another molecule cut by same enzyme. Base pairing occurs. One possible combination 3 DNA ligase seals strands. Recombinant DNA molecule 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 http://get.adobe.com/flashplayer. Methods for making recombinant DNA are central to genetic engineering, the direct manipulation of genes for practical purposes DNA technology has revolutionized biotechnology, the manipulation of organisms or their genetic components to make useful products DNA Technology An example of DNA technology is the microarray, a measurement of gene expression of thousands of different genes. This shows which genes are active in all cells. Another example of DNA technology is Gel Electrophresis Fig. 20-1 Gel Electrophoresis Separates nucleic acids or proteins according to size (density) and charge. DNA cloning To work directly with specific genes, scientists prepare gene-sized pieces of DNA in identical copies, a process called DNA cloning DNA cloning yields multiple copies of a gene or other DNA segments. DNA Cloning and Its Applications: A Preview Most methods for cloning pieces of DNA in the laboratory share general features, such as the use of bacteria and their plasmids Plasmids are small circular extra-chromosomal DNA molecules that replicate separately (autonomously) from the bacterial chromosome. They are often used as vectors in genetic engineering. Cloned genes are useful for making copies of a particular gene and producing a protein product Gene cloning involves using bacteria to make multiple copies of a gene Foreign DNA is inserted into a plasmid, and the recombinant plasmid is inserted into a bacterial cell Reproduction in the bacterial cell results in cloning of the plasmid including the foreign DNA This results in the production of multiple copies of a single gene Cloning a Eukaryotic Gene in a Bacterial Plasmid In gene cloning, the original plasmid is called a cloning vector A cloning vector is a DNA molecule that can carry foreign DNA into a host cell and replicate there 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 http://get.adobe.com/flashplayer. 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 http://get.adobe.com/flashplayer. Amplifying DNA in Vitro: The Polymerase Chain Reaction (PCR) The polymerase chain reaction, PCR, can produce many copies of a specific target segment of DNA know as VNTR’s (specific DNA code repeats unique to an individual). PCR and DNA Replication require the same ingredients to make a new strand (Helicase, DNA polymerase.) A three-step cycle—heating, cooling, and replication—brings about a chain reaction that produces an exponentially growing population of identical DNA molecules Fig. 20-8a 5 TECHNIQUE 3 Target sequence Genomic DNA 3 5 Fig. 20-8b 1 Denaturation 5 3 3 5 2 Annealing Cycle 1 yields 2 molecules Primers 3 Extension New nucleotides Fig. 20-8c Cycle 2 yields 4 molecules Fig. 20-8d Cycle 3 yields 8 molecules; 2 molecules (in white boxes) match target sequence Fig. 20-8 5 TECHNIQUE 3 Target sequence 3 Genomic DNA 1 Denaturation 5 5 3 3 5 2 Annealing Cycle 1 yields 2 molecules Primers 3 Extension New nucleotides Cycle 2 yields 4 molecules Cycle 3 yields 8 molecules; 2 molecules (in white boxes) match target sequence 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 http://get.adobe.com/flashplayer. DNA Fingerprinting Used in criminal investigations: Detectives and forensic scientists use DNA information found at crime scene to understand what transpired and who committed the crime. Everyone’s DNA is unique due to their variable number tandem repeats. Vaccines A vaccine is a substance used to promote the production of antibodies and provide immunity against the pathogen being introduced. Substances is either part of or completely made of a harmless version of the pathogen. DNA vaccines made using one or more genes from the pathogen. DNA vaccines do not have the capability of causing disease.