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Chapter 9. Biotechnology and Genetic Engineering. Biotechnology and Recombinant DNA. Biotechnology Genetic engineering Transgenic organisms (GMOs) “biological factories” Uses recombinant DNA techniques inserts foreign DNA to modify existing DNA sequence. Requires vector

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Chapter 9

Chapter 9

Biotechnology and Genetic Engineering


Biotechnology and recombinant dna
Biotechnology and Recombinant DNA

  • Biotechnology

  • Genetic engineering

    • Transgenic organisms (GMOs)

    • “biological factories”

    • Uses recombinant DNA techniques

      • inserts foreign DNA to modify existing DNA sequence


Figure 9.1.1



Recombinant dna procedures
Recombinant DNA Procedures

  • Protoplast fusion

  • Electroporation

  • Gene gun

  • Microinjection



Electroporation

Microinjection

Gene gun


Restriction enzymes
Restriction enzymes

  • Naturally occurring bacterial enzymes

    • Cut DNA in predictable and controllable manner

    • Generates restriction fragments

      • Covalently joined with ligase

  • Thousands of restriction enzymes

    • Specific target sites

    • Used to excise genes of interest


Gene amplification
Gene Amplification

  • The Polymerase Chain Reaction (PCR)

    • Amplifies small amounts of DNA

    • Creates millions of copies of DNA fragment in matter of hours

    • Many cycles (30-35) of the following steps:

    • Denaturing

    • Annealing

    • Synthesis


Components of pcr reaction
Components of PCR Reaction

  • Template (DNA Fragment)

  • Primers

  • Deoxyribonucleotides

  • DNA polymerase

    • Heat tolerant Taq Polymerase isolated from Thermus aquaticus

  • Cycles are controlled by temperature alterations

    • thermalcycler


Denature

Anneal

Synthesis


  • Gel electrophoresis

    • Separates DNA fragments according to size

      • DNA put into wells in gel

      • Gel subjected to current

      • DNA moves through the gel

        • Fragments separated according to size


Sources of dna
Sources of DNA

1.Gene libraries

  • Extract total DNA from cells and digest with restriction enzymes

  • Create recombinant vectors to generate clones



2. transcriptase Synthetic DNA made with a DNA synthesis machine

  • Exact sequence of gene must be known


Applications of recombinant dna
Applications of Recombinant DNA transcriptase

  • Agricultural

    • Pest and herbicide resistance

    • Improved nutritional value

    • Pharmaceutical proteins added to animal and plant products


  • Medical transcriptase

    • Pharmaceutical proteins

    • Vaccine production

    • Gene therapy


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