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Chapter 9: Biotechnology and Recombinant DNA

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Chapter 9: Biotechnology and Recombinant DNA BIOTECHNOLOGY Use of microorganisms, cells or cell components to make a product Now: being used as – PowerPoint PPT presentation

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Title: Chapter 9: Biotechnology and Recombinant DNA


1
Chapter 9Biotechnology and Recombinant DNA
2
BIOTECHNOLOGY
  • Use of microorganisms, cells or cell components
    to make a product
  • Now being used as "factories" to make chemicals
    not naturally made in the cells
  • Inserting genes......recombinant DNA (rDNA)
    technology
  • Genetic Engineering

3
rDNA Technology
  1. Gene Cloning
  2. Restriction Enzymes

4
Insulin-the First Recombinant Protein
  • Nobel Prizes for insulin
  • 1923 - discovery of insulin patent for insulin
    sold for one dollar
  • 1958 - the first protein with the determined
    amino acid sequence
  • 1969 - the first protein with determined spatial
    conformation (crystallography)
  • Recombinant insulin
  • 1978- human insulin produced in E. coli

5
How to produce human protein in bacteria?
  • What do you need to make a protein?

6
Gene Cloning
  • The goal amplify a given DNA sequence in
    bacterial cells
  • Steps
  • Gene isolation
  • Isolation of vector
  • Manipulation of DNA sequence
  • Cutting -Res
  • Splicing - DNA ligase
  • Transformation of bacteria
  • Selection of correct bacteria

http//www.sumanasinc.com/webcontent/animations/co
ntent/plasmidcloning.html
7
Restriction Endonucleases
  • Enzymes that cleave DNA at a specific sequence
    (usually 4, 6 8bp palindromes)
  • Some leave sticky ends, some leave blunt ends
  • More than 800 are currently available
  • EcoR1

http//highered.mcgraw-hill.com/olcweb/cgi/pluginp
op.cgi?itswf535535/sites/dl/free/0072437316
/120078/bio37.swfRestriction20Endonucleases
8
How do you get the protein out?
  • E. coli and some other bacteria
  • Cells must be broken open and the product
    purified
  • expensive
  • Some bacteria secrete the product
  • Can be removed from the medium
  • Yeast-able to continuously secrete product
  • eukaryotic workhorse of biotechnology

9
Other tools of biotechnology
  • PCR
  • Gel Electrophoresis

10
Polymerase Chain Reaction
  • PCR small samples of DNA are amplified quickly
  • Can make billions of copies in a few hours
  • Thermalcycler set for the desired temp, times,
    and number of cycles
  • Uses DNA pol from thermophilic bacterium
  • Thermus aquaticus
  • Can survive the intense heat
  • http//bcs.whfreeman.com/mga2e/pages/bcs-main.asp?
    s84n003i168vchaptero003004ns0tuid
    0rau0

11
Gel Electrophoresis
  • Used to separate DNA using an electric field
  • Separate based on size
  • http//learn.genetics.utah.edu/content/labs/gel/
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