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Chapter 19- part 2

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  • 19.14 (part 1) A genomic library contains all of the DNA sequences found in an organism’s genome.
  • 19.14 (part 2) A genomic library contains all of the DNA sequences found in an organism’s genome.
  • 19.15a (part 1) A cDNA library contains only those DNA sequences that are transcribed into mRNA.
  • 19.15a (part 2) A cDNA library contains only those DNA sequences that are transcribed into mRNA.
  • 19.15b (part 1) A cDNA library contains only those DNA sequences that are transcribed into mRNA.
  • 19.15b (part 2) A cDNA library contains only those DNA sequences that are transcribed into mRNA.
  • 19.16 Genomic and cDNA libraries can be screened with a probe to find the gene of interest.
  • 19.17a With in situ hybridization, DNA probes are used to determine the cellular or chromosomal location of a gene or its product. (a) A probe with green fluorescence is specific to chromosome 7, revealing a deletion on one copy of chromosome 7. [Addenbrookes Hospital/Photo Researchers.]
  • 19.21 (part 1) The gene for cystic fibrosis was located by positional cloning.
  • 19.21 (part 2) The gene for cystic fibrosis was located by positional cloning.
  • 19.34 (part 1) Transgenic animals have genomes that have been permanently altered through recombinant DNA technology. In the photograph, a mouse embryo is being injected with DNA. [Photograph: Chad Davis/PhotoDisc.]
  • 19.34 (part 2) Transgenic animals have genomes that have been permanently altered through recombinant DNA technology. In the photograph, a mouse embryo is being injected with DNA.
  • 19.35 (part 1) Knockout mice possess a genome in which a gene has been disabled.
  • 19.35 (part 2) Knockout mice possess a genome in which a gene has been disabled.
  • 19.36 The mouse Mus musculus is a model genetic organism. [Chromosome photograph courtesy of Ellen C. Akeson and Muriel T. Davisson, The Jackson Laboratory, Bar Harbor, Maine.]
  • Latter B

Transcript

  • 1. LECTURE CONNECTIONS 19 | Molecular Genetic Analysis and © 2009 W. H. Freeman and Company Biotechnology
  • 2. 19.3 Molecular Techniques Can Be Used to Find Genes of Interest
    • - Shotgun cloning: one first clones a large number of DNA fragments, knowing that one or more contains the DNA of interest.
    • - Gene library: a collection of clones containing all the DNA fragments from one source
      • Creating a genomic DNA library
  • 3.  
  • 4.  
  • 5.
      • - cDNA library: consisting only of those DNA sequences that are transcribed into mRNA
        • Creating a cDNA library
        • Advantages: fragments from actively transcribed genes; and introns do not interrupt the cloned sequence.
        • Disadvantages: contain only sequences that are presence in mature mRNA; and sequences expressed in the tissue from which RNA was isolated.
  • 6.  
  • 7.  
  • 8.  
  • 9.  
  • 10. Gene Libraries
    • Screening DNA libraries
      • Plating clones of the library
      • Probing plated colonies or plaques
  • 11.  
  • 12. In Situ Hybridization
    • DNA probes used to determine the chromosomal location and to visualize a gene while it is in a cell.
    • A labeled probe is applied to the slide.
    • FISH (fluorescence in situ hybridization) is used to identify the chromosomal location of human genes.
    • Determine the tissue distribution of specific mRNA molecules.
  • 13. A) Deletion on one copy of chromosome 7 in humans. B) Presence of mRNA from the tailless gene in Drosophila .
  • 14.
    • Positional Cloning
      • Isolating genes on the basis of their position on a genetic map.
    • In Silico Gene Discovery: high-speed computerized DNA data mining.
    • Application: isolating the gene for cystic fibrosis by positional cloning.
  • 15.  
  • 16.  
  • 17. DNA Fingerprinting (DNA Profiling)
    • DNA fingerprinting: the use of DNA sequences to identify individual persons.
    • Microsatellites: variable number of copies of repeat sequences possessed by many organisms, which can be amplified by PCR.
    • Application: identifying people who died in the collapse of the World Trade Center; identifying criminals; paternity tests; identify specific strains of pathogenic bacteria; etc.
  • 18.  
  • 19. 19.5 Molecular Techniques Are Increasingly Used to Analyze Gene Function
    • Forward genetics:
      • Begins with a phenotype to a gene that encodes the phenotype
    • Reverse genetics:
      • Begins with a gene of unknown function, first inducing mutations and then checking the effect of the mutation on the phenotype
  • 20. 19.5 Molecular Techniques Are Increasingly Used to Analyze Gene Function
    • Site-directed mutagenesis
      • Reverse genetics: creating mutation in particular DNA sequences, and then studying the effects of these mutation on the organisms.
    • Transgenic animals: animals that have been permanently altered by the addition of a DNA sequence to their genome.
  • 21.  
  • 22.  
  • 23. 19.5 Molecular Techniques Are Increasingly Used to Analyze Gene Function
    • Knock out mice: a normal gene of the mouse has been fully disabled.
    • Knock in mice: a mouse carries an inserted DNA sequence at specific locations.
  • 24.  
  • 25.  
  • 26. 19.5 Molecular Techniques Are Increasingly Used to Analyze Gene Function
    • Silencing genes with RNAi
      • Using RNAi for the treatment of human disease: lowering ApoB with RNAi
    • Model Genetic Organism
      • The mouse, Mus musculus
  • 27.  
  • 28. Concept Check 1
      • forward genetics
      • reverse genetics
      • both forward and reverse genetics
      • neither forward nor reverse genetics
    A geneticist is interested in the immune function of mice, and induces random mutations in a number of genes in mice and then determines which of the resulting mutant mice have impaired immune function. This is an example of ________.
  • 29. 19.6 Biotechnology Harnesses the Power of Molecular Genetics
    • Pharmaceuticals
      • Human insulin
      • Specialized bacteria
      • Agricultural products
      • Oligo nucleotide drugs
      • Genetic testing
      • Gene therapy
        • Direct transfer of genes into humans to treat disease