Applications of Recombinant DNA Technology

• Invitro Mutagenesis - What is the effect of changing individual bases in a gene?
  • Mouse globin genes - TATA box
• RFLP - Restriction Fragment Length Polymorphisms
  • Mauriceville and Oak Ridge Neurospora
  • Human analysis for RFLP
• Reverse Genetics - Start with protein and work backwards

  • Genetics	Reverse Genetics
    Mutant Phenotype	Protein Sequence
    Mutant Allele	DNA Sequence
    DNA Sequence	Mutant Allele - Knock Out Technology
    Protein Sequence	Mutant Phenotype

• Expressing Genes in Bacteria
  • T7 polymerase
  • humulin
  • hGH - human growth hormone
  • Mederrhodin A - A -B
• Expressing Genes in Eukaryotes
  • Introduction
  • plasmids for yeast
  • Yeast two ways
  • Yac and modification
  • Agrobacterium tumefaciens
  • t-DNA
  • tobacco - part B
  • t-DNA
  • examples
    • corn borer
    • round up ready
    • Bt cotton
• Drosophila Expressing Beta Galactosidase
• Polysaccharides on Human like Proteins
• Engineering the Mouse - Knock Out or Specific Gene Changes
• Embryonic Stem Cells to Knock Out Mouse - B
• (Rat) Growth Hormone -- Pseudopregnant
  • Big Mouse --
  • Big Salmon

by Kenneth G. Wilson,
Professor of Botany
Miami University
wilsonkg@muohio.edu